Growth promoting peptides and uses thereof

Abstract

A natural peptide comprising a cell growth promoting fragment of a protein selected from the group consisting of SEQUENCE ID NOS: 1 to 13, and a composition comprising a plurality of growth promoting peptides are described. The use of the peptides and compositions in the prevention of skin aging in humans, the treatment of diseases or conditions characterized by damage to epithelial cells or tissues, such as colon cancer and peripheral inflammatory disorders, and in wound treatment are also disclosed. Specific pea and rice protein derived peptides are described in SEQUENCE ID NOS 15 to 505 and 546 to 704.

Description

[0001] Divisional application

[0002] This application is a divisional application of application number 201680053432.6, filed on July 18, 2016, and entitled “Growth-Promoting Peptides and Uses Thereof”. Background Art

[0003] The increasing desire to maintain a youthful appearance has led to a growing interest in new dermatological approaches to treat skin aging, particularly as people live longer and healthier lives. Structural and functional changes have been observed in human skin with aging (Calleja-Agius J. et al., 2013), and these changes are attributed to a variety of factors, such as environmental factors such as ultraviolet radiation from sunlight or internal factors such as hormonal changes during menopause (Affinito P. et al., 1999).

[0004] Alterations in connective tissue in the dermis and epidermis, particularly a reduction in the extracellular matrix, are important contributors to skin wrinkling and sagging, as they induce significant changes in its mechanical properties. Furthermore, the metabolism and synthesis of the extracellular matrix are affected by aging processes, which are controlled by enzyme activities.

[0005] There are currently four main solutions to the anti-aging process:

[0006] Diet and hormone management: Managing our diet with supplements can help combat the aging process. This approach is currently controversial. Some studies suggest that antioxidant supplements, such as vitamin C or lipoic acid, may have anti-aging properties.

[0007] Hormone therapy: A risky and controversial anti-aging solution. People must be extremely cautious with anything involving hormones, as they can have a wide range of side effects. This solution isn't widely used, and most research is still in the animal stage.

[0008] Surgical anti-aging solutions: These are highly effective in the short term, but are expensive and require a long recovery period. Like any surgery, they carry risks and side effects. For example, surgical anti-aging solutions like blepharoplasty and rhytidectomy, where incisions are made at the hairline, carry some risk of bruising, swelling, drooping eyelids, and secondary infection.

[0009] Recently, there has been growing interest in minimally invasive treatments and techniques designed to address issues like wrinkles, facial volume loss, and other skin damage. The most common topical anti-aging solutions are creams and serums. Their active ingredients can be divided into several categories:

[0010] Moisturizing ingredients keep the skin hydrated. They are primarily large polar molecules that bind to water, such as mucopolysaccharides (e.g., hyaluronic acid).

[0011] Collagen and other extracellular matrix-related components help maintain good tissue skin structure by stimulating biosynthesis if the correct receptors are triggered.

[0012] Cell proliferation ingredients are also important because they help promote the regeneration of skin cells, which will synthesize components needed by the skin, such as extracellular matrix and growth factors.

[0013] The object of the present invention is to provide an alternative minimally invasive treatment for skin aging. Summary of the Invention

[0014] The pea genome encodes over 70,000 different proteins. Applicants have identified six of these proteins, each of which contains one or more peptides that promote cell growth and / or proliferation (hereinafter referred to as "growth promoting peptides" or "growth promoting fragments"). Similarly, among the over 60,000 proteins encoded by the rice genome, applicants have identified seven proteins, each of which contains one or more biologically active peptides that generally promote cell growth and / or proliferation. Cell growth promoting fragments of 14 of the identified proteins have been shown to have an effect on elastin and collagen production and cell proliferation ( Figures 1 to 109 ). Specific plant proteins from which natural peptides were derived are provided in SEQUENCE ID NOs: 1-14 and 705 to 716. Specific pea proteins from which peptides were derived include SEQUENCE ID NOs: 1-2 and 7-10, while specific rice proteins from which peptides were derived include SEQUENCE ID NOs: 3-6 and 11-13. Homologs of these proteins are described in SEQUENCE ID NOs: 525 to 564. Specific peptides originally identified in pea proteins are shown in SEQUENCE ID NOs: 15-215 and 283-340. Specific peptides originally identified in rice proteins are shown in SEQUENCE ID NOs: 216-282 and 341-412. Other peptides identified in pea and rice proteins and variants thereof disclosed herein are provided in SEQUENCE ID NOs 418 to 515 and 546 to 704 and 717-775.

[0015] In a first aspect, the present invention provides a peptide that is typically 5 to 50 amino acids in length and comprises (a) a fragment of a pea or rice protein, such as the pea and rice proteins disclosed herein, such as one selected from SEQUENCE ID NOs: 1 to 14 and 705 to 716 and 717-732, or a homolog thereof, or (b) a variant of a fragment, or (c) a fragment of a peptide (hereinafter referred to as a "peptide of the present invention"). In one embodiment, the peptide is biologically active. In one embodiment, the peptide has cell growth or proliferation promoting activity.

[0016] In one embodiment, the peptide of the present invention comprises a sequence selected from the group consisting of SEQUENCE ID NOs: 15-505 and 546-704 and 717-775.

[0017] In one embodiment, the peptide of the invention consists essentially of a sequence selected from the group consisting of SEQUENCE ID NOs: 15-505 and 546-704 and 717-775.

[0018] In one embodiment, the peptide of the present invention consists of about 3-50 amino acids. In one embodiment, the peptide of the present invention consists of about 4-50 amino acids. In one embodiment, the peptide of the present invention consists of about 5-50 amino acids. In one embodiment, the peptide of the present invention consists of about 6-50 amino acids. In one embodiment, the peptide of the present invention consists of about 7-50 amino acids.

[0019] In one embodiment, the fragment has 8 to 37 amino acids.In one embodiment, the fragment has a charge of -10 to +4.

[0020] Preferably, the c-terminal amino acid is not cysteine ​​(C) or methionine (M).

[0021] Preferably, the n-terminal amino acid is not cysteine ​​(C), histidine (H) or proline (P).

[0022] Preferably, the c-terminal domain of the fragment does not contain cysteine ​​(C).

[0023] Preferably, the n-terminal domain of the fragment does not contain cysteine ​​(C).

[0024] Preferably, the fragment does not contain cysteine ​​(C).

[0025] Preferably, the peptide does not contain cysteine ​​(C).

[0026] In one embodiment of the present invention, the peptide comprises a sequence selected from the group consisting of SEQUENCE ID NOs: 15 to 505.

[0027] In one embodiment of the invention, the peptide consists essentially of a sequence selected from the group consisting of SEQUENCE ID NOs: 15-505.

[0028] Preferably, the fragment is selected from SEQUENCE ID NO: 15-215 and 283-340, or a biologically active variant of the fragment.

[0029] Preferably, the fragment is selected from SEQUENCE ID NO: 216-282 and 341-412, or a biologically active variant of the fragment.

[0030] Preferably, the peptide consists of a fragment selected from SEQUENCE ID NO: 15 to 417, or a biologically active variant of a fragment.

[0031] Preferably, the peptide consists of a sequence selected from SEQUENCE ID NO: 15 to 417.

[0032] Preferably, the fragment is selected from SEQUENCE ID NO: 413-417, or a biologically active variant of the fragment.

[0033] In one embodiment, the peptide comprises or consists of a fragment selected from SEQ ID NO: 246, 283, 284, 245 and 42, or a biologically active variant of a fragment.

[0034] In one embodiment, the peptide comprises a fragment selected from SEQ ID NOs: 247 to 268.

[0035] In one embodiment, the peptides of the present invention are modified. In one embodiment, the peptides are modified with protecting groups. In one embodiment, the peptides are modified to increase their lipophilicity. In one embodiment, the peptides are modified to increase their half-life. In one embodiment, the N-terminal or C-terminal amino acid of the peptide is modified. In one embodiment, the N-terminal or C-terminal amino acid of the peptide is modified with protecting groups.

[0036] [SEQUENCE ID NO: 1 (Pea Protein 1 - P13918)]

[0037] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 1, or a homologue thereof, or a biologically active variant of a fragment.

[0038] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 15 to 90, or a biologically active variant of a fragment.

[0039] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which generally comprises a different bioactive fragment of SEQUENCE ID NO: 1 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a first bioactive fragment (or a bioactive variant of a fragment) selected from SEQUENCE ID NOs: 15 to 90, and the second bioactive peptide comprises a second bioactive fragment (or a bioactive variant of a fragment) selected from SEQUENCE ID NOs: 15 to 90.

[0040] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0041] Homologs of pea protein 1 (SEQUENCE ID NO: 1) include Vicia fabia, chickpea (Cicer arietinum), and lentil (Lens culinaris) homologs (SEQ ID NOs: 525 to 527).

[0042] [SEQUENCE ID NO: 2 (Pea Protein 2-Q9M3X6)]

[0043] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 2, or a homologue thereof, or a biologically active variant of a fragment.

[0044] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 91 to 215, or a biologically active variant of a fragment.

[0045] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 2 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 91-215, and the second bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 91-215.

[0046] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0047] Homologs of pea protein 2 (SEQUENCE ID NO: 2) include Pisuma byssinicum, Lathyrus annuus, and Vicia villosa (SEQ IDs 528 to 530).

[0048] [SEQUENCE ID NO: 3 (Rice protein 1 -Q0DEV5)]

[0049] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 3, or a homologue thereof, or a biologically active variant of a fragment.

[0050] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 216-244, or a biologically active variant of a fragment.

[0051] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 3 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 216-244, and the second bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 216-244.

[0052] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0053] Homologs of rice protein 1 (SEQUENCE ID NO: 15) include common wild rice (Oryza rufipogon), medicinal wild rice (Oryza officinalis), and highland barley (Hordeum vulgare subsp. vulgare) (SEQ ID NOs: 531 to 533).

[0054] [SEQUENCE ID NO: 4 (Rice Protein 2-P14323)]

[0055] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 4, or a homologue thereof, or a biologically active variant of a fragment.

[0056] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 245-246, or a biologically active variant of a fragment.

[0057] The present invention also provides a composition comprising at least one or more peptides of the present invention, comprising different biologically active fragments of SEQ ID NO: 4 or homologs thereof. Preferably, the composition comprises a first peptide and a second peptide, wherein the first peptide comprises the biologically active fragment SEQ ID NO: 245 and the second peptide comprises the biologically active fragment SEQ ID NO: 246.

[0058] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0059] Homologs of rice protein 2 (SEQUENCE ID NO: 4) include Oryza brachyantha and Zizania latifolia (SEQ ID NOs: 534 to 536).

[0060] [SEQUENCE ID NO: 5 (Rice Protein 3- P29835)]

[0061] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 5, or a homologue thereof, or a biologically active variant of a fragment.

[0062] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 247-268, or a biologically active variant of a fragment.

[0063] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 5 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 247-268, and the second bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 247-268.

[0064] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0065] Homologs of rice protein 3 (SEQUENCE ID NO: 5) include maize (Zea mays), sorghum (Sorghum bicolor), and millet (Setaria italica) (SEQ IDs 537 to 539).

[0066] [SEQUENCE ID NO: 6 (Rice Protein 4 - P14614)]

[0067] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 6, or a homologue thereof, or a biologically active variant of a fragment.

[0068] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 269-282, or a biologically active variant of a fragment.

[0069] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 6 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 269-282, and the second bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 269-282.

[0070] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0071] Homologs of rice protein 4 (SEQUENCE ID NO: 6) include Oryza sativa Japonica Group and Brochypodium distachyon (SEQ ID NOs: 540 to 542).

[0072] [SEQUENCE ID NO: 7 (pea protein 3-P09918)]

[0073] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 7, or a homologue thereof, or a biologically active variant of a fragment.

[0074] Preferably, the peptide comprises a biologically active fragment of SEQUENCE ID NO: 283, or a biologically active variant of a fragment.

[0075] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 7 or a homolog thereof. Preferably, the composition comprises a bioactive fragment of SEQUENCE ID NO: 283, or a bioactive variant of the fragment.

[0076] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0077] Homologs of pea protein 3 (SEQUENCE ID NO: 7) include Medicago truncatula, Glycine soja, and Phaseolus vulgaris (SEQ ID NOs: 543 to 545).

[0078] [SEQUENCE ID NO: 8 (Pea Protein 4 - P02857)]

[0079] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 8, or a homologue thereof, or a biologically active variant of a fragment.

[0080] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 284-307, or a biologically active variant of a fragment.

[0081] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 8 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 284-307, and the second bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 284-307.

[0082] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0083] Homologs of pea protein 4 (SEQUENCE ID NO: 8) include Vicia sativa, Vicia narbonesis, and Cicer arietinum (SEQ ID NOs: 546-548).

[0084] [SEQUENCE ID NO: 9 (Pea Protein 5 - P02855)]

[0085] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 9, or a homologue thereof, or a biologically active variant of a fragment.

[0086] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 308-339, or a biologically active variant of a fragment.

[0087] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 9 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 308-339, and the second bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 308-339.

[0088] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0089] Homologs of pea protein 5 (SEQUENCE ID NO: 9) include Lathyrus irsutus, Lathyrus cicero, and Lathyrus sativus (SEQ ID NOs: 549 to 551).

[0090] [SEQUENCE ID NO: 10 (Pea Protein 6 - D3VNE1)]

[0091] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 10, or a homologue thereof, or a biologically active variant of a fragment.

[0092] Preferably, the peptide comprises a biologically active fragment of SEQUENCE ID NO: 340, or a biologically active variant of a fragment.

[0093] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 10 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide comprising the bioactive fragment of SEQUENCE ID NO: 340.

[0094] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0095] Homologs of pea protein 6 (SEQUENCE ID NO: 10) include Medicago truncatula, Vicia peregrine, and Vicia lutea (SEQ ID NOs: 552 to 554).

[0096] [SEQUENCE ID NO: 11 (Rice Protein 5 - P07728)]

[0097] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 11, or a homologue thereof, or a biologically active variant of a fragment.

[0098] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 341-358, or a biologically active variant of a fragment.

[0099] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 11 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 341-358, and the second bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 341-358.

[0100] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0101] Homologs of rice protein 5 (SEQUENCE ID NO: 11) include Oryza sativa (Indica Group), Zizania latifolia, and Avena sativa (SEQ ID NOs: 555 to 557).

[0102] [SEQUENCE ID NO: 12 (Rice Protein 6 - P07730)]

[0103] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 12, or a homologue thereof, or a biologically active variant of a fragment.

[0104] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 359-401, or a biologically active variant of a fragment.

[0105] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 12 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 359-401, and the second bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 359-401.

[0106] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0107] Homologs of rice protein 6 (SEQUENCE ID NO: 12) include Oryza brachyantha and Brochypodium distachyon (SEQ ID NOs: 558 to 560).

[0108] [SEQUENCE ID NO: 13 (Rice Protein 7 - Q0D7S0)]

[0109] Preferably, the peptide comprises a biologically active fragment of the protein of SEQUENCE ID NO: 13, or a homologue thereof, or a biologically active variant of a fragment.

[0110] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 402-412, or a biologically active variant of a fragment.

[0111] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 13 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide comprising a bioactive fragment selected from SEQUENCE ID NOs 402-412 and a second bioactive peptide comprising a bioactive fragment selected from SEQUENCE ID NOs 402-412.

[0112] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0113] Homologs of rice protein 7 (SEQUENCE ID NO: 13) include Oryza sativa (Indica Group), Zizania latifolia, and Avena sativa (SEQ ID NOs: 561 to 564).

[0114] [SEQUENCE ID NO: 14 (Staphylococcus aureus protein 1 - P0C1U8)]

[0115] Preferably, the peptide comprises a fragment of the protein of SEQUENCE ID NO: 14, or a homologue thereof.

[0116] Preferably, the peptide comprises a biologically active fragment selected from SEQUENCE ID NOs: 413-417, or a biologically active variant of a fragment.

[0117] The present invention also provides a composition comprising at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, or preferably at least ten bioactive peptides of the present invention, each of which comprises a different bioactive fragment of SEQUENCE ID NO: 14 or a homolog thereof. Preferably, the composition comprises a first bioactive peptide and a second bioactive peptide, wherein the first bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 413-417, and the second bioactive peptide comprises a bioactive fragment selected from SEQUENCE ID NOs 413-417.

[0118] In one embodiment, the biologically active peptide, variant or fragment is a cell growth peptide or a proliferation promoting peptide, variant or fragment, respectively.

[0119] Homologs of Staphylococcus aureus protein 1 (SEQUENCE ID NO: 14) include:

[0120] >gi|580560623|gb|EVF84961.1|Glutamyl endopeptidase [Staphylococcus aureus COAS6020]

[0121] >gi|580687002|gb|EVH10169.1|Glutamyl endopeptidase [Staphylococcus aureus UCIM6080]

[0122] >gi|751815683|gb|KIN24957.l|Glutamyl endopeptidase [Staphylococcus aureus MRSA_CVM43477]

[0123] >gi|781884797|dbj|BAR08486.1|Glutamyl endopeptidase precursor [Staphylococcus aureus subspecies aureus]

[0124] >gi|781887762|dbj|BAR11210.1|Glutamyl endopeptidase precursor [Staphylococcus aureus subsp. aureus]

[0125] The present invention also provides a composition comprising at least one and preferably a plurality of peptides of the present invention, wherein each peptide of the present invention comprises a biologically active fragment of a protein disclosed herein, for example selected from SEQUENCE ID NO: 1 to 14 or a homologue thereof, or a biologically active variant of a fragment.

[0126] Typically, the or each peptide of the invention comprises a biologically active fragment selected from SEQUENCE ID NOs: 15-505 and 546 to 704 and 717-775, or a biologically active variant of a fragment.

[0127] Typically, the or each peptide of the invention is selected from a biologically active fragment selected from SEQUENCE ID NOs: 15-505 and 546 to 704 and 717-775, or a biologically active variant of a fragment.

[0128] The present invention also provides a composition comprising at least one, and preferably a plurality, of the peptides of the present invention, wherein the or each peptide of the present invention comprises a biologically active fragment of a pea or rice protein disclosed herein, which is typically selected from SEQUENCE ID NOs: 1 to 2 and 7 to 10. Typically, the or each peptide of the present invention is selected from or comprises a biologically active fragment selected from SEQUENCE ID NOs: 15 to 215 and 283 to 340, or a biologically active variant of a fragment.

[0129] The present invention also provides a composition comprising at least one and preferably a plurality of peptides of the present invention, wherein the peptide or each peptide of the present invention comprises a biologically active fragment of a protein disclosed herein, which is generally selected from SEQUENCE ID NOs: 3 to 6 and 11 to 13. Typically, the peptide or each peptide of the present invention is selected from or comprises a biologically active fragment selected from SEQUENCE ID NOs: 216 to 282 and 341 to 412, or a biologically active variant of a fragment.

[0130] Preferably, the composition comprises at least two different growth promoting peptides of the invention.

[0131] Preferably, the composition comprises at least three different growth promoting peptides of the invention.

[0132] Preferably, the composition comprises at least four different growth promoting peptides of the invention.

[0133] Preferably, the composition comprises at least five different growth promoting peptides of the invention.

[0134] Preferably, the composition comprises at least six different growth promoting peptides of the invention.

[0135] Preferably, the composition comprises at least seven different growth promoting peptides of the invention.

[0136] Preferably, the composition comprises at least eight different growth promoting peptides of the invention.

[0137] Preferably, the composition comprises at least nine different growth promoting peptides of the invention.

[0138] Preferably, the composition comprises at least ten different growth promoting peptides of the invention.

[0139] In one embodiment, the composition comprises one or more of SEQUENCE ID NOs 247 to 268.

[0140] In one embodiment, the composition comprises one or more of SEQUENCE ID NOs 248, 249, 252, 253, and 257.

[0141] In one embodiment, the invention includes a composition comprising substantially all of the fragments SEQUENCE ID NOs: 15-215 and 283-340, or growth-enhancing variants of the fragments, or mixtures of growth-enhancing fragments and variants.

[0142] In one embodiment, the invention includes a composition comprising substantially all of the fragments SEQUENCE ID NOS: 216-282 and 341-412, or growth-enhancing variants of the fragments, or mixtures of growth-enhancing fragments and variants.

[0143] In one embodiment, the composition is enriched in peptides having a molecular weight of less than 10 kD.

[0144] In one embodiment, the composition is a powder.

[0145] The invention also relates to a plaster, bandage or dressing suitable for application to a wound and comprising a peptide or composition of the invention.

[0146] The present invention also relates to an artificial cell culture medium comprising a peptide of the present invention. The present invention also relates to an artificial cell culture medium comprising a composition of the present invention. In one embodiment, the cell culture medium is formulated for culturing eukaryotic cells. In one embodiment, the cell culture medium is formulated for culturing prokaryotic cells.

[0147] The present invention also relates to a plaster, bandage or dressing suitable for application to a wound and comprising a peptide or composition of the present invention.

[0148] The present invention also relates to a peptide of the present invention for promoting cell growth.

[0149] The present invention also relates to a peptide of the present invention for use in promoting cell culture growth.

[0150] The present invention also relates to a peptide of the present invention for use in promoting tissue growth.

[0151] The present invention also relates to a peptide of the present invention for use in promoting the growth of dermal or epithelial tissue.

[0152] The present invention also relates to a peptide of the present invention for promoting skin growth.

[0153] The present invention also relates to a peptide of the present invention for use in promoting organ growth.

[0154] The present invention also relates to a peptide of the present invention for use in promoting the growth of an organism.

[0155] The present invention also relates to a composition of the present invention for promoting cell growth.

[0156] The present invention also relates to a composition of the present invention for promoting cell culture growth.

[0157] The present invention also relates to a composition of the present invention for promoting tissue growth.

[0158] The present invention also relates to a composition of the present invention for promoting epithelial tissue growth.

[0159] The present invention also relates to a composition of the present invention for promoting skin growth.

[0160] The present invention also relates to a composition of the present invention for promoting organ growth.

[0161] The present invention also relates to a composition according to the invention for use in promoting the growth of an organism.

[0162] In one embodiment, the cell, tissue, or organism has a normal pathology (e.g., aging skin). In one embodiment of the invention, the cell, tissue, or skin has an abnormal pathology (e.g., tissue damaged by trauma, drug use, or epithelial tissue in the gastrointestinal tract damaged by an inflammatory condition).

[0163] Growth promoting uses may be in vivo or in vitro. Growth promoting uses may include external (ie, skin) or internal (ie, gastrointestinal tract) administration to a mammal.

[0164] The present invention also relates to a peptide of the present invention for use in slowing down or inhibiting human skin aging.

[0165] The present invention also relates to a method for slowing or inhibiting the aging of human skin, comprising the step of applying the peptide of the present invention to human skin. Typically, the peptide of the present invention is applied topically to the skin. Administration can be via a plaster or patch or a formulation suitable for topical application.

[0166] The present invention also relates to a composition of the present invention for slowing down or inhibiting human skin aging. The present invention also relates to a peptide of the present invention for preventing or slowing down human skin aging.

[0167] The present invention also relates to a method for slowing down or inhibiting aging of human skin, comprising the step of applying the composition of the present invention to human skin. Typically, the composition of the present invention is topically applied to the skin.

[0168] The present invention also relates to a peptide of the present invention for use in treating wounds in a mammal.

[0169] The present invention also relates to a composition of the peptides of the present invention for use in treating wounds in mammals.

[0170] The present invention also relates to a wound treatment composition or product of the present invention for use in treating a wound in a mammal.

[0171] The present invention also relates to the peptides of the present invention for use in treating or preventing a disease or condition characterized by damaged epithelial cells or tissues.

[0172] The present invention also relates to compositions of the peptides of the present invention for use in treating or preventing a disease or condition characterized by damaged dermal or epithelial cells or tissues.

[0173] In one embodiment, the disease or condition characterized by damaged dermal or epithelial cells or tissue is selected from cancer, trauma.

[0174] The present invention also relates to a peptide of the present invention for use in maintaining or restoring intestinal health in a mammal.

[0175] The present invention also relates to a composition of the peptides of the present invention for use in maintaining or restoring intestinal health in a mammal.

[0176] The present invention also relates to a peptide of the present invention for use in maintaining or restoring muscle health (eg, grams of lean tissue) in a mammal.

[0177] The present invention also relates to a composition of the peptides of the present invention for use in maintaining or restoring muscle health (eg, grams of lean tissue) in a mammal.

[0178] The present invention also relates to a pharmaceutical composition comprising the peptide of the present invention in combination with a pharmaceutically acceptable carrier.

[0179] The present invention also relates to a pharmaceutical composition comprising the peptide composition of the present invention in combination with a pharmaceutically acceptable carrier.

[0180] The present invention also relates to a peptide of the present invention for use in treating or preventing an inflammatory disease in a mammal.

[0181] The present invention also relates to a composition of the present invention for use in treating or preventing an inflammatory disease in a mammal.

[0182] The peptides of the present invention are used in the topical cosmetic or pharmaceutical compositions of the present invention at a cosmetically or pharmaceutically effective concentration to achieve the desired effect; in a preferred form, 0.00000001% (by weight) to 20% (by weight), preferably 0.000001% (by weight) to 15% (by weight), more preferably 0.0001% (by weight) to 10% (by weight), and even more preferably 0.0001% (by weight) to 5% (by weight). Ideally, the peptides of the present invention are preferably used in an amount of about 0.00001% w / w to about 0.5% w / w [0.1 to 5000 ppm] of the composition, more preferably 0.00005 w / w to about 0.05 w / w [0.5 to 500 ppm] of the composition, and most preferably about 0.0001 w / w to about 0.01 w / w [1 to 100 ppm] of the composition. Ideally, the peptides of the present invention are preferably used at about 0.0001% w / w to about 0.004% w / w of the composition.

[0183] For the peptide composition of the present invention, a typical daily dose may be 0.2 g to 100 g.

[0184] The present invention also relates to an artificial personal care composition comprising the peptide of the present invention.

[0185] The present invention also relates to an artificial personal care composition comprising a composition of the peptides of the present invention.

[0186] In one embodiment, the personal care composition is a skin care product. In one embodiment, the personal care composition is a product formulated for topical application to human skin. In one embodiment, the personal care composition is an anti-aging product. In one embodiment, the personal care composition is a toothpaste product. In one embodiment, the personal care composition is a fragrance product. In one embodiment, the personal care composition is a deodorant product. In one embodiment, the personal care composition is an antiperspirant product. In one embodiment, the personal care composition is a soap. In one embodiment, the personal care composition is a liquid soap. In one embodiment, the personal care composition is a cream. In one embodiment, the personal care composition is a lotion. In one embodiment, the personal care composition is a gel. In one embodiment, the personal care composition is a powder.

[0187] The present invention also relates to an artificial wound treatment composition comprising a peptide of the present invention. The present invention also relates to an artificial wound treatment composition comprising a composition of the present invention. Typically, wound treatment compositions are formulated for topical application to wounds. In one embodiment, the composition comprises a cream, gel, lotion, or powder.

[0188] The present invention also provides topical compositions comprising peptides of the present invention. It should be understood that topical compositions can comprise a variety of peptides, fragments and / or variants. In one embodiment, topical compositions comprise all peptides substantially. In one embodiment, topical compositions comprise all variants substantially. Topical compositions of the present invention can be present in preparations selected from the group comprising creams, composite emulsions, anhydrous compositions, aqueous dispersions, oils, breasts, balms, foams, lotions, gels, curds, hydroalcoholic solutions, aqueous glycolic acid solutions, cosmetics, personal care products, hydrogels, liniments, sera, soaps, dusting powders, pastes, semisolid preparations, liniments, clear solutions, shampoos, conditioners, ointments, any rinses, talc, mousses, powders, sprays, aerosols, solutions, suspensions, emulsions, syrups, elixirs, polysaccharide films, patches, gel patches, bandages, adhesive systems, water-in-oil emulsions, oil-in-water emulsions and silicone emulsions.

[0189] In one embodiment of the present invention, the emulsion contains a lipid or oil. The emulsion can be, but is not limited to, an oil-in-water, water-in-oil, water-in-oil-in-water, and oil-in-water-in-silcone emulsions. The emulsion can contain a moisturizer. The emulsion can contain a defoaming agent such as silicone. The emulsion can have any suitable viscosity. The emulsion can also contain an emulsifier and / or a defoaming agent. Methods for preparing emulsions are known to those skilled in the art.

[0190] Topical composition of the present invention can be incorporated into the medical device for administration.This apparatus can include but is not limited to fabric, patch, bandage, meter, socks, tights, underwear, dressing, gloves, face mask, adhesive patch, non-adhesive patch, occlusive patch and micro-electric patch or suitable adhesive system.In such embodiment, apparatus and stratum corneum (such as skin) directly contact, thereby discharge peptide of the present invention.Should be appreciated that topical composition can be incorporated in any suitable form that this paper describes in detail.For example, topical composition of the present invention or peptide can be incorporated in device or be present on the device surface or can be any suitable preparation of emulsifier, gel or wax preparation or this paper definition and be incorporated in device or on the surface of device.Apparatus can be suitable for adhering to or being attached to on skin.

[0191] In one embodiment, the device is adapted to release a constant amount of a composition or peptide of the invention. It will be appreciated that the amount of composition contained in the sustained-release system will depend, for example, on where the composition will be administered, the kinetics and duration of release of the composition of the invention, and the nature of the condition, disorder, and / or disease to be treated and / or cared for. The device may be such that the composition is released by biodegradation of the device due to body moisture, skin pH, or body temperature, or by friction between the device and the body.

[0192] In one embodiment of the present invention, the topical composition may also include at least one cosmetic or pharmaceutically acceptable excipient. Excipients may be used interchangeably with functional ingredients or additives. It should be understood that although the topical compositions of the present invention may be applied alone, they are typically applied in admixture with cosmetic or pharmaceutical excipients. Cosmetically or pharmaceutically acceptable excipients are well known in the art and are any known excipient that can be used, as long as they are suitable for topical application and are dermatologically acceptable without abnormal toxicity, incompatibility, and / or allergic reactions.

[0193] Preferably, any excipients included are present in trace amounts. The amount of excipient included will depend on many factors, including the type of excipient used, the nature of the excipient, the ingredients of the topical composition, the amount of active ingredient or peptide in the topical composition, and / or the intended use of the topical composition. The nature and amount of any excipient should not unacceptably alter the benefits of the peptides of the present invention.

[0194] In one embodiment of the present invention, the excipient may be a suitable diluent, carrier, binder, lubricant, suspending agent, coating agent, preservative, stabilizer, dye, carrier, solubilizer, base, emollient, emulsifier, fragrance, humectant and / or surfactant.

[0195] Examples of suitable diluents include, but are not limited to, any of the diluents disclosed in US2014120131 or US2004132667. Examples include ethanol, glycerol, and water.

[0196] Examples of suitable carriers include, but are not limited to, lactose, starch, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol, and any suitable carrier disclosed in US2014120131 or US2004132667.

[0197] Examples of suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose, anhydrous lactose, free-flowing lactose, beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose and polyethylene glycol, and any suitable binder disclosed in US2014120131 or US2004132667.

[0198] Examples of suitable lubricants include, but are not limited to, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride, and any suitable lubricant disclosed in US2014120131 or US2004132667.

[0199] Carrier can be any suitable carrier known in the art or disclosed in US2014120131 or US2004132667. In some embodiments, carrier can include but is not limited to liquid, such as water, oil or surfactant, including those derived from petroleum, animals, plants or synthesis, polymer, oil (such as peanut oil, mineral oil, castor oil, soybean oil), alcohol, polysorbate, sorbitan ester, ether sulfate (ether sulfates), sulfate, betaine, glycoside, maltoside (maltosides), fatty alcohol, nonoxynol ether, poloxamer, polyoxyethylene, polyethylene glycol, dextrose, glycerol or digitonin. It should be understood that carrier will be dermatologically acceptable. Preferred carrier contains emulsion, such as oil-in-water, water-in-oil, water-in-oil-in-water and silicone-in-water-in-oil emulsion. Emulsion can also contain emulsifier and / or defoamer.

[0200] In one embodiment of the present invention, the topical composition may further comprise one or more additional ingredients. The topical composition of the present invention may be applied continuously, simultaneously, or sequentially with one or more other additional agents. Such additional ingredients may be those that would be beneficial to include in the topical composition, or would be beneficial depending on the intended use of the topical composition. The additional ingredients may be active or functional or both active and functional.

[0201] Examples of such additional ingredients include, but are not limited to, one or more of cleansers, conditioners, sunscreens, pigments, moisturizers, thickeners, gelling agents, essential oils, astringents, pigments, anticoagulants, antifoaming agents, binders, additives, buffers, chelating agents, topical analgesics, film formers or materials, fillers, polymers, opacifiers, pH adjusters, propellants, reducing agents, sequestrants, skin whitening and lightening agents, skin conditioning agents, aloe vera, healing agents, soothing agents, slip agents, pantothenic acid, treatment agents, thickeners, vitamins, colorants, pharmaceuticals, preservatives, antifoaming agents, buffers, astringents, polymers, pH adjusters, deodorants, or any other dermatologically acceptable carrier or surfactant.

[0202] It should be understood that the additional ingredients listed may provide more than one benefit. The classifications given herein are for clarity and convenience only and are not intended to limit the additional ingredients to the specific applications or categories listed.

[0203] Any additional ingredients should be suitable for application to the skin without undue toxicity, incompatibility and / or allergic reaction.

[0204] In some embodiments, the additional ingredients have glucose transport activity or contribute to glucose transport activity. In some embodiments, the additional ingredients have anti-inflammatory activity or contribute to anti-inflammatory activity. In some embodiments, the additional ingredients have anti-aging activity or contribute to anti-aging activity. In some embodiments, the additional ingredients are used for stratum corneum health and / or development, skin health and / or development, and / or muscle health, recovery and / or development. The activating agent can be a pharmacological enhancer. Such activating agents are known and available on the market. In this case, the topical composition of the present invention can be applied continuously, simultaneously or one after another with one or more other activating agents.

[0205] In some embodiments, the additional ingredients may be farnesol ([2E,6E],-3,7,11,-trimethyl-2,6,10,dodecatrien-1-ol), phytantriol (3,7,11,15,tetramethylhexadecane-1,2,3,-triol), desquamation actives, enzymes, enzyme inhibitors, enzyme activators, plant and marine extracts, anti-acne actives, anti-wrinkle or anti-atrophy actives, antioxidants / free radical scavengers, chelating agents, flavonoids, anti-inflammatory agents, anti-cellulite agents, local anesthetics, tanning actives. actives), skin lightening agents, skin repair agents, bisabolol, antimicrobial or antifungal actives, sunscreen actives, particulate matter, conditioning agents, structuring agents, thickeners.

[0206] The desquamation active may be any suitable agent that enhances skin appearance or skin texture and as disclosed in US2014120131 or US2004132667.

[0207] Examples of anti-acne actives are disclosed in US2014120131 or US2004132667 and include resorcinol, salicylic acid, erythromycin, zinc, sulfur, benzoyl peroxide.

[0208] Examples of the thickener are disclosed in US2014120131 or US2004132667, and include carboxylic acid polymers, cross-linked polyacrylate polymers, polyacrylamide polymers, polysaccharides.

[0209] Examples of conditioning agents are disclosed in US2014120131 or US2004132667, and include moisturizers, emollients, or skin conditioning agents.

[0210] Examples of structuring agents are disclosed in US2014120131 or US2004132667 and include any agent that provides rheological properties to the composition and contributes to the stability of the composition.

[0211] Any suitable antimicrobial or antifungal active can be used, and examples include those disclosed in US2014120131 or US2004132667. These actives are capable of destroying microorganisms, preventing their growth or action. Examples include, but are not limited to, beta-lactams, quinolones, tetracyclines, erythromycin, streptomycin sulfate, salicylic acid, and benzoyl peroxide.

[0212] Examples of particulate materials include metal oxides. Examples of anti-cellulite agents include xanthines. Examples of tanning actives include 1,3-dihydroxy-2-propanone and those disclosed in US2014120131 or US2004132667. Examples of local anesthetics include benzocaine, lidocaine, and bupivacaine and those disclosed in US2014120131 or US2004132667.

[0213] Examples of skin lightening agents include any agents known in the art, such as kojic acid, ascorbic acid, and those disclosed in US2014120131 or US2004132667.

[0214] Examples of sunscreen actives include any suitable organic or inorganic sunscreen actives. Examples include metal oxides, 2-ethylhexyl-p-methoxycinnamate, and those disclosed in US2014120131 or US2004132667.

[0215] Examples of skin repair agents include pantothenic acid as disclosed in US2014120131 or US2004132667.

[0216] Examples of anti-inflammatory agents include any agent that enhances the appearance, skin tone, or color of the skin, including but not limited to corticosteroids, hydrocortisone, nonsteroidal agents such as ibuprofen and aspirin, and those disclosed in US2014120131 or US2004132667.

[0217] Examples of flavonoids include flavanones, methoxyflavones, unsubstituted chalcone, and mixtures thereof, as well as those disclosed in US2014120131 or US2004132667.

[0218] The example of enzyme includes lipase, protease, catalase, superoxide dismutase, amylase, peroxidase, glucuronidase, ceramidase, hyaluronidase.The example of enzyme inhibitor includes trypsin inhibitor, Bowman Birk inhibitor (Bowmann Birk inhibitors), chymotrypsin inhibitor, plant extract, flavonoid, quercetin chalcone and those disclosed in US2014120131 or US2004132667 and its mixture.The example of enzyme activator includes coenzyme A, Q10 (ubiquinone), glycyrrhizin, berberine, chrysin and those disclosed in US2014120131 or US2004132667 and its mixture.

[0219] Examples of anti-wrinkle or anti-atrophy actives include sulfur-containing D and L amino acids, particularly N-acyl derivatives such as N-acetyl-L-cysteine, hydroxy acids, phytic acid, lipoic acid, lysophosphatidic acid, skin peeling agents, vitamin B3, retinoids and those disclosed in US2014120131 or US2004132667, and mixtures thereof.

[0220] The antioxidant / free radical scavenger can be any agent useful for providing protection against UV radiation or other environmental agents that may cause skin damage, such as those disclosed in US2014120131 or US2004132667. Examples of antioxidants / free radical scavengers include ascorbic acid, its salts and derivatives (vitamin C), tocopherol and its salts and derivatives (vitamin E), butylated hydroxybenzoic acid and its salts, peroxides, gallic acid and alkyl esters, sorbic acid, lipoic acid, amines, lycine pidolate, arginine pilolate, nordihydroguaiaretic acid, bioflavonoids, curcumin, lysine, methionine, proline, superoxide dismutase, silymarin, tea extract, and mixtures thereof.

[0221] Examples of chelating agents include EDTA, NTA, hydroxamic acid, phytic acid, lactoferrin, and those disclosed in US2014120131 or US2004132667, and mixtures thereof. Chelating agents are agents that remove metal ions by forming complexes, rendering them unable to participate in or catalyze chemical reactions. Chelating agents can be used to protect against ultraviolet radiation or other environmental factors that may cause skin damage.

[0222] It should be understood that a variety of additional ingredients can be added. The amount of the additional ingredient can be from about 0.001 to about 50% by weight of the composition, preferably from about 0.01% to about 20%, preferably from about 0.1% to about 10%, from about 0.5% to about 10%, from about 1% to about 5%, preferably 2% by weight of the composition. The amount of the additional ingredient included will depend on many factors, including the type of additional ingredient used, the nature of the additional ingredient, the ingredients of the topical composition, the amount of active substance or peptide in the topical composition and / or the intended use of the topical composition. The nature and amount of any additional ingredient should not unacceptably alter the benefits of the peptides of the present invention.

[0223] The topical compositions may be alcohol-free.

[0224] In some embodiments of the present invention, except the peptide of the present invention (also referred to as the active substance of composition), composition also comprises one or more additional activating agents.In addition or alternatively, composition can be used together with one or more other additional activating agents.Usually the additional activating agent is only present in trace amounts.In some embodiments, there may not be an additional activating agent in the composition.The amount of the additional activating agent comprised will depend on many factors, including the type of additional activating agent used, the character of the additional activating agent, the composition of the topical composition, the active substance in the topical composition or the amount of peptide and / or the intended use of the topical composition.The character and amount of any additional activating agent should not unacceptably change the benefit of the peptide of the present invention.

[0225] It should be understood that an ingredient considered an "active" ingredient in one product may be a "functional" or "excipient" ingredient in another, and vice versa. It should also be understood that some ingredients play a dual role as both an active ingredient and a functional or excipient ingredient.

[0226] Examples of additional active agents include glucose transport promoting drugs, skin supplements, skin treatment and / or care agents, anti-inflammatory agents, anti-aging agents, cell growth promoters, and pharmacological enhancers. Such agents are well known in the art, and it will be appreciated that any suitable additional active agent may be used. Additional active agents for treating and / or caring for the skin may include collagen synthesis agents, retinoids, exfoliants, fat reducing agents, elastase inhibitors, melanin synthesis stimulators or inhibitors, self-tanning agents, anti-aging agents, antimicrobials, antifungals, antifungal agents, bactericides, and repair agents. Active agents also include anti-inflammatory agents.

[0227] Any additional active agent should be suitable for application to the skin without undue toxicity, incompatibility and / or allergic response.

[0228] It should be understood that the classifications given herein are for clarity and convenience only and are not intended to limit the additional ingredients, excipients, or actives to the particular applications or categories listed.

[0229] In a particularly preferred embodiment, the methods and uses of the present invention include administering a peptide or composition of the present invention in combination with one or more other active agents (e.g., existing growth promoting drugs or pharmacological enhancers available on the market). In this case, the composition of the present invention can be administered continuously, simultaneously or sequentially with the one or more other active agents.

[0230] The effects of the present invention are achieved by topical application or administration of the topical compositions of the present invention as described herein to a human, animal, or patient in need of treatment or care. Topical delivery preferably refers to delivery to the stratum corneum, such as the skin, hair, and / or nails, but may also refer to delivery to body cavities lined with epithelial cells, such as the lungs or airways, gastrointestinal tract, buccal cavity. The effects may be localized to the surface of the skin or may be within the skin, or both.

[0231] The topical composition of the present invention is applied in an amount that is cosmetically or pharmaceutically effective. In other words, in an amount that is nontoxic but sufficient to provide the desired effect. It should be understood that those skilled in the art will be able to determine the appropriate dose for applying the topical composition of the present invention without excessive experimentation. Alternatively, the physician will determine the actual dose that is most suitable for the patient based on the specific condition to be treated or nursed, the disease or illness, and the age, weight, and / or health status of the individual. This will depend on a variety of factors, including the activity of the specific compound employed, the metabolic stability and duration of action of the compound, age, weight, general health, sex, diet, mode of application and time, excretion rate, drug combination, the severity of the specific condition, and the individual being treated. Certainly, individual cases (wherein higher or lower dosage ranges are suitable) can exist, and these are also within the scope of the present invention. For example, the composition can be applied in a dosage of 0.01 to 50 mg / kg body weight, such as 0.1 to 30 mg / kg, more preferably 0.1 to 20 mg / kg body weight, more preferably 0.1 to 10 mg / kg body weight, preferably 0.1 to 5 mg / kg body weight. In an exemplary embodiment, one or more doses of 10 to 300 mg / day, or more preferably 10 to 150 mg / day, are administered to the patient. The dosage and frequency are best suited to the purpose. The frequency of application or administration can vary widely depending on the needs of each subject, and the recommended application or administration ranges from once a month to ten times a day, preferably once a week to four times a day, more preferably from three times a week to three times a day, and even more preferably once or twice a day.

[0232] In a preferred embodiment, repeated use of the topical composition is provided.

[0233] Can be by but not limited to friction or massage into keratinous tissue to be treated or nursing, skin or body area to apply topical composition.In some embodiments, composition stays on the area of ​​health or is not removed from the area of ​​health.In other embodiments, after a period of time, such as but not limited to about 2 minutes to 60 minutes, about 5 minutes to about 30 minutes, preferably about 10 minutes to about 20 minutes after removing composition.Can remove composition immediately after application.In some embodiments of the present invention, composition of the present invention can be applied to area to be treated by making composition of the present invention and / or peptide obtain greater penetration means, such as but not limited to iontophoresis, ultrasound electrophoresis, electroporation, micro-electric patch, mechanical pressure, osmotic pressure gradient, occlusive solidification, by means of microinjection or needleless injection of pressure, such as by oxygen pressure injection, or its any combination.

[0234] The peptides of the present invention are used in the topical cosmetic or pharmaceutical compositions of the present invention in a cosmetically or pharmaceutically effective concentration to achieve the desired effect; in a preferred form with respect to the total weight of the composition, it is 0.00000001% (by weight) to 20% (by weight); preferably 0.000001% (by weight) to 15% (by weight), more preferably 0.0001% (by weight) to 10% (by weight), and even more preferably 0.0001% (by weight) to 5% (by weight).

[0235] In some embodiments of the present invention, the composition can be delivered via any of liposomes, mixed liposomes, oleosomes, niosomes, ethosomes, microcapsules, capsules, macrocapsules, nanocapsules, nanostructured lipid carriers, sponges, cyclodextrins, vesicles, micelles, mixed micelles of surfactants, mixed micelles of surfactants and phospholipids, microspheres, spheres, lipid spheres, particles, nanospheres, nanoparticles, microparticles, solid nanoparticles, and microemulsions (including water-in-oil microemulsions and nanoemulsion microspheres and microparticles having an internal structure of reverse micelles).

[0236] A variety of methods can be used to prepare liposomes. See, e.g., Szoka et al., Ann. Rev. Biophys. Bioeng. 9:467 (1980), U.S. Pat. Nos. 4,186,183, 4,217,344, 4,235,871, 4,261,975, 4,485,054, 4,501,728, 4,774,085, 4,837,028, 4,235,871, 4,261,975, 4,485,054, 4,501,728, 4,774,085, 4,837,028, 4,946,787, PCT Publication No. WO 91 / 17424, Deamer & Bangham, Biochim. Biophys. Acta 443:629-634 (1976); Fraley et al., PNAS 76:3348-3352 (1979); Hope et al., Biochim. Biophys. Acta 812:55-65 (1985); Mayer et al., Biochim. Biophys. Acta 858:161-168 (1986); Williams et al., PNAS 85:242-246 (1988); Liposomes (Ostro (ed.), 1983, Chapter 1); Hope et al., Chem. Phys. Lip. 40:89 (1986); Gregoriadis, Liposome Technology (1984) and Lasic, Liposomes: from Physics to Applications (1993)). Suitable methods include, for example, sonication, extrusion, high pressure / homogenization, microfluidization, detergent dialysis, calcium-induced fusion of small liposome carriers, and ether fusion methods, all of which are well known in the art.

[0237] These delivery systems can be suitable for achieving greater penetration of the compounds of the present invention and / or peptides. This can improve pharmacokinetic and pharmacodynamic properties. The delivery system can be a sustained-release system, in which the compounds of the present invention or peptides are gradually released over a period of time and preferably have a constant release rate over a period of time. Delivery systems are prepared by methods known in the art. The amount of the peptide contained in the sustained-release system will depend on where the composition will be delivered and the duration of release and the type of the condition, disease and / or illness to be treated or cared for.

[0238] The topical compositions of the present invention may be administered to humans or animals for either human or veterinary use.

[0239] The topical compositions of the present invention may be used for pharmaceutical, personal care and / or cosmetic purposes.

[0240] The compositions can be used to treat or care for any disease, disorder or condition of the skin, including but not limited to psoriasis, dermatitis, atopic dermatitis, eczema, spongioma, edema, skin cancer, ulcers, acne, scars, cellulitis, elastosis, keratosis, rosacea, varicose veins, inflammatory diseases.

[0241] Topical compositions can be used to treat or manage visible signs of aging, including but not limited to wrinkles, stretch marks and dark circles, dryness, fine lines, age spots, erythema, sagging skin, and conditions caused by sun exposure (including sunburn), stress, pollution, and / or diet. Topical compositions can also be used to delay, slow, or inhibit the onset of skin or aging. The compositions can be administered via a medical device, such as a plaster or patch as described herein.

[0242] The topical composition can be used to treat or care for wounds in mammals. In another embodiment, the topical composition is used to treat or prevent a disease or condition characterized by damaged epithelial cells or tissues, and / or damaged dermal or epithelial cells or tissues. The disease can be, but is not limited to, cancer and trauma.

[0243] The topical compositions can be used to treat or care for any muscle condition, improve muscle condition in a mammal, promote muscle recovery, typically after exercise, maintain or restore muscle health (e.g., grams of lean tissue) in a mammal, enhance physical performance, treat or prevent a disease or condition characterized by lethargy or low energy.

[0244] The topical compositions can be used to promote tissue growth, promote epithelial tissue growth, promote skin growth, promote organ growth, promote organism growth.The skin can have normal pathology and / or abnormal pathology.

[0245] The topical compositions may also be used to treat or care for any inflammatory condition.

[0246] Other aspects of the present invention relate to a pharmaceutical composition comprising a peptide of the present invention or a composition of the peptide of the present invention mixed with one or more pharmaceutically acceptable diluents, excipients or carriers. Although the peptides and compositions of the present invention can be administered alone, they are typically administered in admixture with a pharmaceutical carrier, excipient or diluent, particularly for human treatment. The pharmaceutical composition can be used for humans or animals for human or veterinary purposes. Examples of such suitable excipients for the various forms of pharmaceutical compositions described herein can be found in "Handbook of Pharmaceutical Excipients, 2nd Edition, ed., by A. Wade and P. J. Weller (1994)". ndIn particular, formulations for topical delivery are described in Topical drug delivery formulations, edited by David Osborne and Antonio Aman, Taylor & Francis, the entire contents of which are incorporated herein by reference. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (AR Gennaro ed., 1985). edit.1985)). Examples of suitable carriers include lactose, starch, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol, and the like. Examples of suitable diluents include ethanol, glycerol, and water. The choice of pharmaceutical carrier, excipient, or diluent can be selected according to the intended route of administration and standard pharmaceutical practice. The pharmaceutical composition may contain, for example, or in addition to the carrier, excipient, or diluent, any suitable binder, lubricant, suspending agent, coating agent, solubilizer. Examples of suitable binders include starch, gelatin, natural sugars (such as glucose, anhydrous lactose, free-flowing lactose, β-lactose, corn sweeteners), natural and synthetic gums (such as gum arabic), tragacanth gum or sodium alginate, carboxymethyl cellulose, and polyethylene glycol. Examples of suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like.

[0247] Preservatives, stabilizers, dyes, and even flavorings may be provided in the pharmaceutical composition. Examples of preservatives include sodium benzoate, sorbic acid, and esters of p-hydroxybenzoic acid. Antioxidants and suspending agents may also be used.

[0248] The peptide or composition of the present invention can be suitable for topical, oral, rectal, parenteral, intramuscular, intraperitoneal, intraarterial, intrabronchial, subcutaneous, intradermal, intravenous, nasal, vaginal, buccal or sublingual administration. For oral administration, compressed tablets, pills, tablets, gels, drops and capsules are particularly used. Preferably, these compositions contain 1-250 mg per dose, and more preferably 10-100 mg of active ingredient. Other forms of administration include solutions or emulsions that can be injected intravenously, intraarterially, subcutaneously, intradermally, intraperitoneally or intramuscularly, and the solutions or emulsions are prepared from sterile or sterilizable solutions. The pharmaceutical composition of the present invention can also be in the form of suppositories, vaginal rings, pessaries, suspensions, emulsions, lotions, ointments, creams, gels, sprays, solutions or dusting powders. The composition of the present invention can be formulated for local delivery. Topical delivery generally means delivery to the skin, but can also mean delivery to a body cavity lined with epithelial cells, such as the lungs or airways, gastrointestinal tract, buccal cavity. In particular, preparations for topical delivery are described in Topical drug delivery formulations compiled by David Osborne and Antonio Aman, Taylor & Francis, the entire contents of which are incorporated herein by reference. Compositions or preparations for delivery to the airways are described in (Respiratory Care, 2002, November 47) (Respir Care, 2002, Nov. 47), EP2050437, WO2005023290, US2010098660 and US20070053845 by O'Riordan et al. Compositions and preparations for delivering active agents to the ileum, particularly the proximal ileum, include microparticles and microcapsules, wherein the active agent is encapsulated in a protective matrix formed by polymers or milk proteins, which is acid-resistant but easily dissolves in the more alkaline environment of the ileum. The example of this delivery system is described in EP1072600.2 and EP13171757.1. An alternative method for transdermal administration is by using a skin patch. For example, the active ingredient can be incorporated into a cream consisting of an aqueous emulsion of polyethylene glycol or liquid paraffin. The active ingredient can also be incorporated into an ointment consisting of a white wax or white soft paraffin matrix and such stabilizers and preservatives that may be needed at a concentration of 1 to 10 wt.%.

[0249] Injectable forms may contain 10-1000 mg, preferably 10-250 mg, of active ingredient per dose.

[0250] The compositions may be formulated in unit dosage form, ie, in discrete portions containing a unit dose or multiples or subunits of a unit dose.

[0251] Those of ordinary skill in the art can easily determine the appropriate dosage of one of the compositions of the present invention to be administered to a subject without excessive experimentation. Typically, the physician will determine the actual dosage that is most suitable for the individual patient, and this will depend on a variety of factors, including the activity of the specific compound used, the metabolic stability and duration of action of the compound, age, body weight, general health, sex, diet, mode of administration and time, excretion rate, drug combination, the severity of a particular condition, and the individual being treated. The dosage disclosed herein is an example of an average case. Of course, there can be individual cases (wherein higher or lower dosage ranges are suitable), and these are also within the scope of the present invention. As needed, 0.01 to 30 mg / kg body weight, such as 0.1 to 10 mg / kg, more preferably 0.1 to 1 mg / kg body weight dosage administration medicament. In an exemplary embodiment, in order to treat inflammatory diseases, 10 to 300 mg / day, or more preferably 10 to 150 mg / day one or more dosages will be administered to the patient.

[0252] In a particularly preferred embodiment, the methods and uses of the present invention include administering a peptide or composition of the present invention in combination with one or more other active agents (e.g., existing anti-inflammatory drugs or pharmacological enhancers available on the market). In this case, the composition of the present invention can be administered continuously, simultaneously or sequentially with one or more other active agents.

[0253] In one embodiment of the present invention, the peptide of the present invention can be administered in the form of a conjugate comprising the peptide, and the peptide of the present invention can optionally include a linker and a partner molecule, such as a protein, such as an antibody molecule intended to increase the half-life of the conjugate in vivo. In one embodiment, the peptide can be modified to replace one or more amino acids with amino acids for connecting to partner molecules. For example, in order to conjugate to partner molecules, such as PEG molecules, amino acids can be replaced with lysine residues.

[0254] definition

[0255] All publications, patents, patent applications, and other references mentioned herein are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference and were set forth in its entirety.

[0256] As used herein, unless expressly stated otherwise, the following terms are intended to have the following meanings, in addition to any broader (or narrower) meanings that such terms may have in the art:

[0257] Unless the context requires otherwise, the use of the singular herein should be understood to include the plural, and vice versa. The use of the term "a" or "an" with respect to an entity should be understood to refer to one or more of that entity. Thus, the terms "a" (or "an"), "one or more," and "at least one" are used interchangeably herein.

[0258] As used herein, the term "comprise" or variations thereof, such as "comprises" or "comprising" should be understood to indicate the inclusion of any stated integer (e.g., feature, element, characteristic, property, method / process step, or limitation) or group of integers (e.g., feature, element, characteristic, property, method / process step, or limitation), but not the exclusion of any other integer or group of integers. Thus, as used herein, the term "comprising" is inclusive or open-ended and does not exclude additional, unrecited integers or method / process steps.

[0259] As used herein, the term "disease" is used to define any abnormal condition that impairs physiological function and is associated with specific symptoms. The term is used broadly to encompass any disorder, illness, abnormality, pathology, sickness, condition, or syndrome in which physiological function is impaired, regardless of the nature of the cause (or whether the etiological basis of the disease is actually determined). Thus, it encompasses conditions caused by infection, trauma, injury, surgery, radiation ablation, poisoning, or nutritional deficiencies.

[0260] As used herein, the term "treatment" or "treating" refers to an intervention (e.g., administering an agent to a subject) that cures, ameliorates, or alleviates the symptoms of a disease or eliminates its cause (or alleviates the effects of its cause) (e.g., reducing the accumulation of pathological levels of lysosomal enzymes). In this context, the term is used synonymously with the term "therapy."

[0261] In addition, the term "treatment" or "treating" refers to an intervention (e.g., administering an agent to a subject) that prevents or delays the onset or progression of a disease, or reduces (or eradicates) its incidence in the treated population. In this context, the term treatment is used synonymously with the term "prophylaxis."

[0262] As used herein, an effective amount or therapeutically effective amount of a medicament defines an amount that can be administered to a subject without excessive toxicity, irritation, allergic reaction or other problems or complications, commensurate with a reasonable benefit / risk ratio, but sufficient to provide the desired effect, such as a treatment or prevention that is manifested as a permanent or temporary improvement in the subject's condition. The dosage varies depending on the subject, depending on the age and overall condition of the individual, the mode of administration and other factors. Therefore, although it is impossible to specify an exact effective amount, those skilled in the art will be able to determine the appropriate "effective" amount in any individual case using routine experiments and background common sense. The therapeutic outcome in this case includes eradication or alleviation of symptoms, relief of pain or discomfort, prolonged survival, improvement in mobility and other signs of clinical improvement. The therapeutic outcome is not necessarily a complete cure.

[0263] The term "human or animal" should be understood to mean a human or a mammal or a non-mammal (such as fish).

[0264] The term "composition" should be understood to mean a composition made by the hand of man and not a naturally occurring substance. Exemplary compositions include pharmaceutical compositions and personal care compositions.

[0265] Term used herein " peptide " refers to the polymer being made up of 5 to 50 amino acid monomers that are usually connected by peptide bonds.As the present invention and for the peptide of the present invention (including its fragment and variant), can be wholly or partly by chemical synthesis or by producing from nucleic acid expression.For example, can be according to the standard liquid of sufficient confirmation known in the art, or preferably solid phase peptide synthesis method easily prepares the peptide of the present invention and for the peptide of the present invention (see, for example, JM Stewart and J.D. Young, Solid Phase Peptide Synthesis (2nd edition, Pierce Chemical Company, Rockford, Illinois (1994)) (JM Stewart and J.D. Young, Solid Phase Peptide Synthesis, 2nd edition, Pierce Chemical Company, Rockford, Illinois (1984)); Peptide synthesis in M.Bodanzsky and A.Bodanzsky, Springer Press, New York (1984) (M.Bodanzsky and A.Bodanzsky, The Practice of Peptide Synthesis, Springer Verlag, New York York (1984)). If necessary, any peptide used in the present invention may be chemically modified to increase their stability. Chemically modified peptides or peptide analogs include any functional chemical equivalent of a peptide characterized by improved stability and / or efficacy in vivo or in vitro relative to the practice of the present invention. The term peptide analog also refers to any amino acid derivative of a peptide as described herein. Peptide analogs can be prepared by procedures including, but not limited to, modification of side chains, incorporation of non-natural amino acids and / or derivatives thereof during peptide synthesis, and the use of cross-linking agents and other methods of imposing conformational constraints on peptides or their analogs. Examples of side chain modifications include modification of amino groups, such as by reaction with an aldehyde followed by reductive alkylation with NaBH4; Amidation with methylacetimidate; acetylation with acetic anhydride; carbamoylation of amino groups with cyanates; trinitrobenzylation of amino groups with 2,4,6-trinitrobenzylic acid (TNBS); alkylation of amino groups with succinic anhydride and tetrahydrophthalic anhydride; and pyridoxylation of lysine with pyridoxal-5'-phosphate followed by reduction with NABH4. The guanidinium group of arginine residues can be modified by forming heterocyclic condensation products with reagents such as 2,3-butanedione, phenylglyoxal, and glyoxal.Carboxyl groups can be modified by carbodiimide activation via O-acylisourea formation, followed by subsequent derivatization (e.g., to the corresponding amide). Sulfhydryl groups can be modified by methods such as carboxymethylation with iodoacetic acid or iodoacetamide; oxidation with performic acid to form cysteic acid; formation of mixed disulfides with other thiol compounds; reaction with maleimide, maleic anhydride, or other substituted maleimides; formation of mercury derivatives using 4-chloromercuric benzoic acid, 4-chloromercuric benzenesulfonic acid, phenylmercuric chloride, 2-chloromercuric-4-nitrophenol, and other mercurials; and carbamoylation with cyanate esters at alkaline pH. Tryptophan residues can be modified by, for example, oxidation with N-bromosuccinimide or alkylation of the indole ring with 2-hydroxy-5-nitrobenzyl bromide or sulfonyl halides. Tyrosine residues can be modified by nitration with tetranitromethane to form 3-nitrotyrosine derivatives. Modification of the imidazole ring of the histidine residue can be accomplished by alkylation with iodoacetic acid derivatives or N-carbethoxylation with diethyl pyrocarbonate. Examples of incorporation of non-natural amino acids and derivatives during peptide synthesis include, but are not limited to, the use of norleucine, 4-aminobutyric acid, 4-amino-3-hydroxy-5-phenylpentanoic acid, 6-aminohexanoic acid, tert-butylglycine, norvaline, phenylglycine, ornithine, sarcosine, 4-amino-3-hydroxy-6-methylheptanoic acid, 2-thienylalanine and / or D-isomers of amino acids. Peptide structural modifications include the generation of retro-inverso peptides comprising reverse sequences encoded by D-amino acids.

[0266] The term "modified peptide" is used interchangeably with the term derivative of a peptide. Modified peptides include peptides that have been replaced by one or more groups as defined herein. The modification can be any modification that provides a peptide and / or composition of the present invention with an increased ability to penetrate cells. The modification can be any modification that increases the half-life of the composition or peptide of the present invention. In one embodiment, the group is a protecting group. The protecting group can be an N-terminal protecting group, a C-terminal protecting group or a side chain protecting group. The peptide can have one or more of these protecting groups. Those skilled in the art know suitable techniques for reacting amino acids with these protecting groups. These groups can be added by preparation methods known in the art, such as the methods outlined in paragraphs

[0104] to

[0107] of US2014120141. These groups can remain on the peptide or may be removed. Protective groups can be added during the synthesis process. In one embodiment of the invention, the peptide can be substituted by a group selected from one or more straight or branched, long or short chain, saturated or unsaturated, substituted by hydroxyl, amino, aminoacyl, sulfate or sulfide groups or unsubstituted groups with 1 to 29 carbon atoms. N-acyl derivatives include those derived from acetic acid, capric acid, lauric acid, myristic acid, caprylic acid, palmitic acid, stearic acid, behenic acid, linoleic acid, linolenic acid, lipoic acid, oleic acid, isostearic acid, elaidoic acid, 2-ethylhexaneic acid, coconut oil fatty acid, tallow fatty acid, hardened tallow fatty acid, palm kernel fatty acid, lanolin fatty acid or similar acids. These can be substituted or unsubstituted. When substituted, they are preferably substituted by hydroxyl or sulfur-containing groups, such as but not limited to SO3H, SH or SS. In one embodiment of the invention, the peptide is R1-X-R2. The R1 and / or R2 groups are bound to the amino terminus (N-terminus) and carboxyl terminus (C-terminus) of the peptide sequence, respectively. In one embodiment, the peptide is R1-X. Alternatively, the peptide is X-R2. Preferably, R1 is H, C 1-4alkyl, acetyl, benzoyl or trifluoroacetyl; X is a peptide of the present invention; R2 is OH or NH2. In one embodiment, R1 is selected from the group formed by H, acyclic substituted or unsubstituted aliphatic groups, substituted or unsubstituted alicyclic groups, substituted or unsubstituted heterocyclic groups, substituted or unsubstituted heteroarylalkyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted arylalkyl groups, tert-butoxycarbonyl groups, 9-fluorenylmethoxycarbonyl groups (Fmoc) and R5-CO-, wherein R5 is selected from the group formed by H, acyclic substituted or unsubstituted aliphatic groups, substituted or unsubstituted alicyclic groups, substituted or unsubstituted aryl groups, substituted or unsubstituted unsubstituted aralkyl, substituted or unsubstituted heterocyclic, and substituted or unsubstituted heteroarylalkyl; R2 is selected from the group consisting of -NR3R4, -OR3 and -SR3, wherein R3 and R4 are independently selected from the group consisting of H, acyclic substituted or unsubstituted aliphatic, substituted or unsubstituted alicyclic, substituted or unsubstituted heterocyclic, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl; and with the proviso that R1 and R2 are not α-amino acids. According to another preferred embodiment, R2 is -NR3R4, -OR3 or -SR3, wherein R3 and R4 are independently selected from the group consisting of H, substituted or unsubstituted C1-C 24 Alkyl, substituted or unsubstituted C2-C 24 alkenyl, tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl (Fmoc), substituted or unsubstituted C2-C 24 Alkynyl, substituted or unsubstituted C3-C 24 Cycloalkyl, substituted or unsubstituted C5-C 24 Cycloalkenyl, substituted or unsubstituted C8-C 24 Cycloalkynyl, substituted or unsubstituted C6-C 30 Aryl, substituted or unsubstituted C7-C 24 The group consisting of arylalkyl, 3-10 substituted or unsubstituted heterocyclyl ring, and 2-24 carbon atoms and 1-3 atoms (excluding carbon), wherein the alkyl chain has 1-6 carbon atoms. Optionally, R3 and R4 can be bonded to the nitrogen atom through a saturated or unsaturated carbon-carbon bond to form a ring. More preferably, R2 is -NR3R4 or -OR3, wherein R3 and R4 are independently selected from H, substituted or unsubstituted C1-C 24 Alkyl, substituted or unsubstituted C2-C 24 Alkenyl, substituted or unsubstituted C2-C 24 Alkynyl, substituted or unsubstituted C3-C 10 Cycloalkyl, substituted or unsubstituted C6-C 15The invention further comprises the following: an alkyl radical, ...

[0267] According to another embodiment of the present invention, R1 is selected from the group consisting of H, acetyl, lauroyl, myristoyl or palmitoyl, and R2 is -NR3R4 or -OR3, wherein R3 and R4 are independently selected from H, methyl, ethyl, hexyl, dodecyl and hexadecyl, preferably R2 is -OH or -NH2. More preferably, R1 is acetyl or palmitoyl, and R2 is -NH2. In a preferred embodiment, the acyl group is bonded to the N-terminus of at least one amino acid of the peptide. In one embodiment of the present invention, the peptide is modified to include a side chain protecting group. The side chain protecting group can be one or more of a benzyl or benzyl-based group, a tert-butyl-based group, a benzyloxy-carbonyl (Z) group and an allyloxycarbonyl (allyloxycarbonyl) (allyloxycarbonyl (alloc)) protecting group. The side chain protecting group can be derived from an achiral amino acid, such as achiral glycine. The use of an achiral amino acid helps to stabilize the resulting peptide and is also conducive to the simple synthetic route of the present invention. Preferably, the peptide further comprises a modified C-terminus, preferably an amidated C-terminus. The achiral residue may be α-aminoisobutyric acid (methylalaine). It will be appreciated that the specific side chain protecting group used will depend on the sequence of the peptide and the type of N-terminal protecting group used.

[0268] "Conjugate": In one embodiment of the invention, the peptide is conjugated, linked or fused to a binding partner, such as one or more polyethylene glycol polymers or other compounds, such as molecular weight-increasing compounds or lipophilic groups. The molecular weight-increasing compound is any compound that increases the molecular weight of the resulting conjugate, typically by 10% to 90%, or 20% to 50%, and can have a molecular weight of 200 to 20,000, preferably 500 to 10,000. The molecular weight-increasing compound can be PEG, any water-soluble (amphiphilic or hydrophilic) polymer moiety, a homopolymer or copolymer of PEG, a monomethyl-substituted polymer of PEG (mPEG) and polyoxyethylene glycerol (POG), polyamino acids (such as polylysine, polyglutamic acid, polyaspartic acid), particularly those in the L configuration, pharmacologically inactive proteins (such as albumin), gelatin, fatty acids, polysaccharides, lipid amino acids and dextran. The polymer portion may be linear or branched, and may have a molecular weight of 500 to 40,000 Da, 5,000 to 10,000 Da, or 10,000 to 5,000 Da. The compound (binding partner) may be any suitable cell penetrating compound, such as a tat peptide, a penetratin, or a pep-1 peptide. The compound (binding partner) may be an antibody molecule.

[0269] The compound (binding partner) can be a lipophilic portion or a polymer portion. Lipophilic substituents and polymer substituents are known in the art. Lipophilic substituents include acyl groups, sulfonyl groups, N atoms, O atoms or S atoms that form part of an ester, sulfonyl ester, thioester, amide or sulfonamide. The lipophilic portion can include a hydrocarbon chain with 4 to 30 C atoms, preferably 8 to 12 C atoms. It can be linear or branched, saturated or unsaturated. The hydrocarbon chain can be further substituted. It can be a cycloalkane or a heterocycloalkane. The peptide can be modified at the N-terminus, C-terminus or both. The polymer or compound (binding partner) is preferably connected to an amino group, a carboxyl group or a sulfhydryl group, and can be connected through the N-terminus or C-terminus of the side chain of any amino acid residue. The polymer or compound (binding partner) can be conjugated to the side chain of any suitable residue. The polymer or compound (binding partner) can be conjugated through a spacer. The spacer can be a natural or non-natural amino acid, succinic acid, lysyl, glutamyl, aspartyl, glycyl, β-alanyl, γ-aminobutyryl. The polymer or compound (binding partner) can be conjugated through an ester, sulfonyl ester, thioester, amide, carbamate, urea, or sulfonamide. Those skilled in the art will appreciate suitable methods for preparing the conjugate.

[0270] "Fragment" means a fragment of a protein selected from SEQUENCE ID NOs: 1 to 14, the fragment typically being 7 to 37 consecutive amino acids in length and typically having a charge of -9 to +3; the c-terminal amino acid typically being not cysteine ​​(C) or methionine (M), and the n-terminal amino acid typically being not cysteine ​​(C), histidine (H), proline (P), or threonine (T). The charge of a peptide, fragment, or region is determined using the method of Cameselle, JC, Ribeiro, JM, and Sillero, A. (1986). The formula is derived and used to calculate the net charge of an acid-base compound. It is applicable to amino acids, proteins, and nucleotides, see Biochem. Educ. 14, 131-136.

[0271] The term "natural" as applied to a peptide means a peptide that comprises (a) a fragment of a plant protein, typically rice or pea protein, or a variant of pea protein (including lentil, sugar snap pea, or chickpea), or a variant of rice protein (including oat, grass, corn, wild rice, and banana), or (b) a variant of a fragment of a plant protein, such as a fragment of a homolog of a plant protein. The peptides or fragments of the present invention can be isolated from plant proteins or synthetically prepared using methods known to those skilled in the art and described herein.

[0272] "C-terminal domain" as applied to a fragment means the first three amino acids at the c-terminus of the fragment.

[0273] "N-terminal domain" as applied to a fragment means the last three amino acids at the n-terminus of the fragment.

[0274] "Biological activity" as applied to a peptide or fragment means having a health promoting effect when administered to a mammal, such as one or more of glucose transport promotion, antibacterial, anti-inflammatory, or promotion of cell growth or proliferation. In one embodiment, the term "biological activity" means cell growth promotion.

[0275] "Growth promoting" or "growth promoting activity" as applied to a peptide or fragment means that the peptide or fragment is capable of increasing elastin production or cell proliferation in human skin treated with a 20 μM solution of the peptide or fragment described in the assay below.

[0276] "Glucose transport promotion" or "glucose transport promoting activity" applied to a peptide, variant or fragment means that when used at a concentration of 2 μM in the following in vitro assay, it is a peptide, variant or fragment that can increase GLUT4 translocation to skeletal muscle compared to an untreated control. L6-GLUT4myc cells were grown in 10% FBS and 2 μg / ml of blasticidin. Cells were grown for 48-72 hours before being seeded at 15,000 cells per well in 2% FBS in a 24-well plate and differentiated for 6 to 8 days prior to the experiment. L6-GLUT4myc cells were serum starved for 3 hours before being incubated with 100 nM of insulin for 30 minutes, or with 200, 20, 2.0 and 0.2 μM of SP and 2, 1, 0.5 and 0.25 mg / ml of peptide / peptide compositions for 3 hours. Based on previous findings, a 3-hour incubation period was selected to determine that incubation with branched-chain amino acids containing dipeptides for 3 hours increased glucose uptake in L6 myotubes 1. In order to determine the GLUT4myc translocation at the same time point, the treatment was staggered. The amount of myc-tagged GLUT4 on the cell surface was measured by antibody-coupled colorimetric assay. Briefly, after incubation with insulin for 30 minutes or synthetic peptides or peptide compositions for 3 hours, L6-GLUT4myc cells were fixed by incubation with 3% paraformaldehyde (PFA). Then, 0.1M glycine solution was added to quench PFA, and cells were blocked with 5% goat serum. Myotube monolayers were exposed to anti-myc antibodies and then incubated with peroxidase-conjugated donkey anti-mouse IgG. 1mL of o-phenylenediamine dihydrochloride (OPD) reagent was added to each well, and the reaction was terminated by adding 250μl / well of 3M HCl. To determine GLUT4 translocation to the cell surface, aliquots of each condition were measured spectrophotometrically on a microplate reader using absorbance at 492 nm. Preferably, the peptide or fragment is capable of increasing GLUT4 translocation by at least 50% (i.e., an increase in relative units of GLUT4 translocation of 1% to 1.5%) compared to an untreated control.

[0277] "Antibacterial" or "antibacterial activity" as applied to a peptide or fragment means a peptide or fragment that is able to visibly inhibit bacterial growth in the following agar plate-based growth inhibition assay: Peptide stock solution dissolved in DMSO = 5 mg / mL. The bacterial inoculum was adjusted to a McFarland 0.5 standard and the MHA plates were wiped. A blank plate was placed in the plate and 10 μL of each compound (at 64 μg / mL - the maximum tested concentration) was added. The plates were incubated at 37°C for 16-18 hours. Appropriate controls (DMSO; hydrolyzed casein (Mueller-Hinton) medium alone; and two antibiotic tablets - ciprofloxacin and tetracycline) were also performed.

[0278] "Anti-inflammatory" as applied to a peptide or fragment means that when macrophages are treated with 100 μM of the peptide or fragment, the peptide or fragment can significantly reduce the secretion of TNFα by LPS-stimulated J774.2 macrophages (compared to untreated LPS-stimulated J774.2 macrophages). J774.2 macrophages were treated with 100 μM of the synthetic peptide for 24 hours and then stimulated with (A) LPS (10 ng / ml) for 5 hours, or (B) LPS (10 ng / ml) for 5 hours and then with ATP (5 mM) for 1 hour. The supernatant was collected and the level of TNFα was determined by ELISA.

[0279] "Enriched in peptides having a molecular weight of less than 10KD" as applied to the compositions of the present invention means that the dry weight percentage of peptides in the composition having a molecular weight of less than 10KD is greater than the dry weight percentage of polypeptides / proteins in the composition having a molecular weight of 10KD or more.

[0280] "Homologs" of a reference protein should be understood to mean proteins from different plant species that have at least 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity with the reference protein. Thus, for example, homologs of pea protein P13918 include:

[0281] >gi|137584|sp|P08438.1|VCL_VICFA RecName: Intact = Vicilin; Flag: Precursor [Vicia faba]

[0282] >gi|22057|emb|CAA68559.1|Via Globulin [Vicia faba var. minor]>gi|383931031|gb|AFH56916.1|Via Globulin [Vicia faba var. minor]

[0283] >gi|502105533|ref|XP_004492829.1|Predicted: Vicilin-like isoform X1 [Cicer arietinum] Chickpea (ChickPea)

[0284] >gi|29539109|emb|CAD87730.1|Allergen Len c 1.0101 [Lens culinaris] Lentils

[0285] A "variant" of a growth promoting fragment should be taken to mean a fragment having substantially the same amino acid sequence as a reference growth promoting fragment and which has growth promoting activity as defined above. Thus, for example, the term should be taken to include fragments in which one or more amino acid residues are altered. Preferably, such alterations involve insertions, additions, deletions and / or substitutions of only 5 or fewer amino acids, more preferably 4 or fewer, even more preferably 3 or fewer, most preferably 1 or 2 amino acids. Insertions, additions and substitutions with natural and modified amino acids are envisaged. The variant may have conservative amino acid changes, wherein the introduced amino acid is structurally, chemically or functionally similar to the amino acid being replaced. Typically, the variant has at least 70% amino acid sequence homology to the reference growth promoting fragment, preferably at least 80% sequence homology, more preferably at least 90% sequence homology, and ideally at least 95%, 96%, 97%, 98% or 99% sequence homology.

[0286] In this specification, the term "sequence identity" should be understood to include both sequence identity and similarity, that is, a variant (or homolog) that shares 70% sequence identity with a reference sequence is a variant (or homolog) in which any 70% of the aligned residues of the variant (or homolog) are identical or conservatively substituted with the corresponding residues in the reference sequence over the entire length of the sequence. Sequence identity is the amount of the trait that completely matches between two different sequences. Therefore, gaps are not counted, and the measurement is related to the shorter of the two sequences. With regard to "sequence homology", the term should be understood to mean that when the percentage of aligned residues of the variant (or homolog) is identical or conservatively substituted with the corresponding residues in the reference sequence and in the case where the variant (or homolog) shares the same function with the reference sequence, the variant (or homolog) shares a defined percentage similarity or identity with the reference sequence. This comparison and percentage homology or sequence identity can be determined using software programs known in the art, for example, one alignment program is BLAST using default parameters. The detailed information of these programs can be learned at the following Internet address: http: / / www.ncbi.nlm.nih.gov / blast / Blast.cgi .

[0287] Variants of SEQUENCE ID NO:448 (QSFLLSGNQ)

[0288] Provided below are variants of SEQUENCE ID NO: 448 (QSFLLSGNQ), including variants having 1 or 2 conservative amino acid substitutions, 1, 2 to 3 non-conservative amino acid substitutions, 1-2 amino acid additions, 1, 2 or 3 amino acid deletions.

[0289] A conservative amino acid substitution:

[0290] QSFILSGNE, ESFLLSGNQ, QSYLLSGNQ, QSFLLSGDQ (SEQ ID NO 418-421)

[0291] Two conservative amino acid substitutions:

[0292] QSYLLSGNE, ESFLLSGNE, ESYLLSGNQ, QSFLLSGDE, QSYLLSGDQ (SEQ ID NO 422 to 426)

[0293] A non-conservative amino acid substitution

[0294] QSFRLSGNQ, QSFLLSYNQ, QFFLLSGNQ, QSFLLSGAQ, QSFLLSGNP (SEQ ID NO 427 to 431)

[0295] Two non-conservative amino acid substitutions

[0296] QSFRRRSGNQ, QSFLLSYIQ, QFFLLSGNL, ​​QSFLLSGAQ, QSFLLSGNP (SEQ ID NO 432 to 436)

[0297] One or two amino acids added

[0298] QSFLLSGNQQ, QSFLLLSGNQ, AQSFGLLSGNQ, RQSFLLISGNQ, QSFLLSGNQK (SEQ ID NO437 to 441)

[0299] One, two, or three amino acid deletions

[0300] QFLLSGNQ, SFLLSGNQ, QSFLLSGN, QSFLLGNQ, QSFLSGNQ, QSLLSGNQ, SFLLSGNQ, QSFLLSGN, SFLLSGN, QSFSGNQ (SEQ ID NO 442 to 451)

[0301] The term "variant" also includes fragments of the peptides of the present invention. "Fragment of a peptide of the present invention" or "peptide fragment" means a fragment of one of the peptides of the present invention having at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acids and generally having biological activity, such as anti-inflammatory activity, anti-aging activity, glucose transport promoting activity or antibacterial activity. In one embodiment, the fragment consists of at least 30%, 40%, 50%, 60%, 70%, 80% or 90% of the reference sequence. Therefore, the present invention also provides biologically active fragments of the peptides of the present invention, as well as peptides comprising one or more of these fragments. In one embodiment, the fragment is biologically active. In one embodiment, the fragment is a cell growth or proliferation promoting fragment. Examples of fragments of the peptides of the present invention are provided in SEQUENCE ID NOs 494 to 524.

[0302] "Anti-aging" means inhibiting or slowing the appearance of aging in human skin and / or reversing the appearance of aging. "Slowing or inhibiting skin aging" means slowing or inhibiting the aging process in the skin and / or reversing the appearance of aging.

[0303] By "disease or condition characterized by damage to dermal or epithelial cells or tissues" is meant any condition or disease that results in damage to dermal or epithelial tissues or cells or organs. One example is trauma, which typically results in damage to the skin. Another example is an inflammatory skin condition, such as psoriasis or eczema, which typically results in damage to the skin. Another example is an inflammatory disorder of the small intestine, which may result in damage to the epithelial cells / tissue lining the small intestine. Another example is damage to the epithelial cells / tissue lining the small intestine due to the ingestion of toxic or noxious substances (e.g., toxic chemicals or drugs). Another example is cancer, such as bowel cancer, which may result in damage to the epithelial tissue in the intestine. Another condition is a peripheral inflammatory disorder, such as atopic dermatitis, which can result in damage to the skin in humans.

[0304] "Inflammatory conditions" means immune-mediated inflammatory conditions that affect humans and are generally characterized by abnormal expression (dysregulated expression) of one or more cytokines. Examples of inflammatory conditions include skin inflammatory conditions, arthritis inflammatory conditions, inflammatory conditions of the cardiovascular system, certain autoimmune diseases, lung and airway inflammatory conditions, and intestinal inflammatory conditions. Examples of skin inflammatory conditions include dermatitis, such as atopic dermatitis and contact dermatitis, acne vulgaris, and psoriasis. Examples of arthritis inflammatory conditions include rheumatoid arthritis. Examples of cardiovascular inflammatory diseases are cardiovascular disease and atherosclerosis. Examples of autoimmune diseases include type 1 diabetes, Graves' disease, Guillain-Barré disease, lupus, psoriatic arthritis, and ulcerative colitis. Examples of lung and airway inflammatory conditions include asthma, cystic fibrosis, COPD, emphysema, and acute respiratory distress syndrome. Examples of intestinal inflammatory conditions include colitis and inflammatory bowel disease. Other inflammatory conditions include cancer, hay fever, periodontitis, allergies, hypersensitivity reactions, ischemia, depression, systemic diseases, post-infectious inflammation and bronchitis.The present invention also relates to a peptide or composition of the present invention for use in treating an inflammatory condition in a mammal.

[0305] "Metabolic disorders" should be understood to include prediabetes, diabetes; type 1 diabetes; type 2 diabetes; metabolic syndrome; obesity; diabetic dyslipidemia; hyperlipidemia; hypertension; hypertriglyceridemia; hyperfattyacidemia; hypercholerterolemia; hyperinsulinemia and MODY. The present invention also relates to a peptide or composition of the present invention for use in treating a metabolic disorder in a mammal.

[0306] By "disease or condition characterized by damage to dermal or epithelial cells or tissues" is meant any condition or disease that results in damage to dermal or epithelial tissues or cells or organs. One example is trauma, which typically results in damage to the skin. Another example is an inflammatory skin condition, such as psoriasis or eczema, which typically results in damage to the skin. Another example is an inflammatory disorder of the small intestine, which may result in damage to the epithelial cells / tissue lining the small intestine. Another example is damage to the epithelial cells / tissue lining the small intestine due to the ingestion of toxic or noxious substances (e.g., toxic chemicals or drugs). Another example is cancer, such as bowel cancer, which may result in damage to the epithelial tissue in the intestine. Another condition is a peripheral inflammatory disorder, such as atopic dermatitis, which can result in damage to the skin in humans.

[0307] "Disease or condition characterized by bacterial infection" means any condition or disease characterized by pathology caused by bacterial growth or infection, including, for example, MRSA, Salmonella, Listeria, bacterial pneumonia, staphylococcal food poisoning, bacterial meningitis. Specific examples are provided at https: / / en.wikipedia.org / wiki / List of infectious diseases.

[0308] "Maintaining or restoring gut health" means reducing and / or regulating the pro-inflammatory response in the gut, and more specifically the epithelial cells. A healthy microbiome provides some protection against pathogenic viruses and bacteria, and their presence is required to guide the development of our immune system. It has been shown that these bacteria can respond to the body's signals of stress, illness, or age, which can manifest through inflammation, and therefore turn on their pathogenic genes and cause or contribute to disease. With the ability to be reduced and maintained at healthy levels, the inflammatory response can help maintain healthy bacteria. Digestive problems comprise the number one health issue in North America and seem to be appearing more frequently in recent years. One way to maintain digestive health is to maintain appropriate inflammation and gut flora.

[0309] "Improving muscle status" means improving muscle health, such as promoting skeletal muscle protein synthesis, bone glucose uptake, improving grams of lean tissue in therapeutic or non-therapeutic situations, generally promoting muscle recovery after active exercise, or improving muscle performance. The method or use can be therapeutic or non-therapeutic. The term "improving lean tissue status" should be understood to mean increasing lean tissue grams, or inhibiting or preventing the rate of decrease in lean tissue grams.

[0310] By "promoting muscle recovery" is meant that glucose uptake in skeletal muscle is increased compared to untreated skeletal muscle.

[0311] "Disease or condition characterized by lethargy or low energy" means any condition or disease characterized by feeling tired or having low energy. Examples include allergies, asthma, anemia, cancer and its treatment, chronic pain, heart disease, infection, depression, eating disorders, grief, sleep disorders, thyroid problems, medication side effects, alcohol use, or drug use.

[0312] "Maintaining or restoring muscle health" means helping to maintain or restore muscle health in mammals caused by damage incurred during exercise. By promoting glucose transport in skeletal muscle, peptides promote post-exercise recovery and reduce muscle soreness / pain and damage associated with exercise. They can also be used to reduce and prevent muscle cramps and allow rapid recovery from muscle cramps. Cramps can be caused by physical stress, psychological stress and / or repetitive strain injury stress. By promoting glucose transport, peptides help reduce muscle myopathy and help prevent sarcopenia in mammals, promote recovery from damage during exercise, and reduce muscle soreness / exercise-related pain and damage. The present invention also relates to a peptide or composition of the present invention for maintaining or restoring muscle health in mammals.

[0313] In the present specification, the term "substantially all" as applied to a list of peptides should be understood to mean at least 60%, 70%, 80%, 90% or 95% of the peptides. BRIEF DESCRIPTION OF THE DRAWINGS

[0314] Figures 1 to 100 : Effects of the synthetic peptides of the present invention on the proliferation of human dermal fibroblasts (HDF).

[0315] Figure 101 : Effects of the synthetic peptide of the present invention (SEQ ID 42) on elastin synthesis in human dermal fibroblasts (HDF).

[0316] Figure 102 : Effects of the synthetic peptide of the present invention (SEQ ID 42) on collagen synthesis in human dermal fibroblasts (HDF).

[0317] Figure 103 : Effects of the synthetic peptide of the present invention (SEQ ID 701) on elastin synthesis in human dermal fibroblasts (HDF).

[0318] Figure 104 : Effects of the synthetic peptide of the present invention (SEQ ID 701) on collagen synthesis in human dermal fibroblasts (HDF).

[0319] Figure 105 : Effects of the synthetic peptide of the present invention (SEQ ID 246) on elastin synthesis in human dermal fibroblasts (HDF).

[0320] Figure 106 : Effects of the synthetic peptide of the present invention (SEQ ID 246) on collagen synthesis in human dermal fibroblasts (HDFs).

[0321] Figure 107: Effects of the synthetic peptide of the present invention (SEQ ID 284) on elastin synthesis in human dermal fibroblasts (HDFs).

[0322] Figure 108 : Effects of the synthetic peptide of the present invention (SEQ ID 245) on elastin synthesis in human dermal fibroblasts (HDF).

[0323] Figure 109 : Effects of the synthetic peptide of the present invention (SEQ ID 245) on collagen synthesis in human dermal fibroblasts (HDF).

[0324] Figure 110 .Shows the integrity control and feasibility control of the measurement system.

[0325] Figure 111 For peptides P1, P2, and P3, the percentage of elastin expression in the superficial dermis compared to the control group (water or DMSO)

[0326] * It shows a significant increase in elastin expression in the superficial and middle dermis.

[0327] Figure 112 For peptides P1, P2, and P3, the percentage of elastin expression in the mid-dermis compared to the control group (water or DMSO)

[0328] * Significant increase in elastin expression was observed in the superficial and mid-dermis.

[0329] Figure 113 Percentage of cell proliferation in the epidermal basal layer for peptides P6 and P8 and peptide combinations P9 and P10 compared to control (water or DMSO)

[0330] * Indicates a significant increase.

[0331] Figure 114 . Histological analysis of elastic fibers (+catechin, x200).

[0332] Figure 115 . Immunohistochemical evaluation of mitotic index (Ki67, x400). DETAILED DESCRIPTION

[0333] Example 1 - Cell proliferation assay

[0334] BrDu is incorporated into newly synthesized DNA chains in actively proliferating cells.

[0335] Following partial denaturation of double-stranded DNA, immunochemical detection of BrdU allows assessment of the cell population that is synthesizing DNA.

[0336] Human dermal fibroblasts (HDF-Sigma 10605a) were seeded at 10,000 cells / well in a 96-well plate in DMEM containing 10% fetal calf serum (FCS), 1% penicillin / streptomycin (Pen / strep), and 1% L-glutamine and maintained therefor for 24 hours.

[0337] After the initial 24 h incubation, cells were incubated with 5 μg / ml, 0.5 μg / ml or 0.05 μg / ml of the synthetic peptide for 24 h.

[0338] After incubation with the synthetic peptide for 18 hours, 20 μL of BrDu reagent was added to each well.

[0339] During the 24-hour incubation, cells were fixed and the amount of 2-DG6P was measured using a BrdU cell proliferation assay. All steps were performed according to the manufacturer's instructions.

[0340] Results are calculated as a percentage of the untreated control. An increase in optical density readings indicates greater incorporation of BrDU and increased cell proliferation.

[0341] The results are shown in Figure 1-100 and Table 1 below.

[0342] Table 1

[0343]

[0344]

[0345] Example 2 - Collagen production assay

[0346] Hydroxyproline in tissue preparations is a direct measure of the amount of collagen present. The FIRELISA Human Hydroxyproline ELISA kit assay is designed to measure hydroxyproline in tissue or peptide compositions.

[0347] Human dermal fibroblasts (HDF-Sigma 10605a) were seeded at 50,000 cells / well in a 24-well plate in DMEM containing 10% fetal calf serum (FCS), 1% penicillin / streptomycin (Pen / strep), and 1% L-glutamine and maintained for 24 hours.

[0348] After the initial 24 hour incubation, the cells were incubated with 5 μg / ml, 1 μg / ml or 0.1 μg / ml of synthetic peptide for 96 hours respectively.

[0349] After treatment, cells were lysed using 4 freeze-thaw cycles in liquid nitrogen. Lysed cells were centrifuged and 50 μl / ml of each supernatant was assayed using the FIRELISA Human Hydroxyproline ELISA kit. All steps were performed according to the manufacturer's instructions.

[0350] Results were calculated as a percentage of the untreated control group. An increase in the optical density reading indicates an increase in collagen content.

[0351] Results are shown in Figure 102 , 104 , 106 and 109.

[0352] Example 3 - Elastin Production Assay

[0353] Elastin is a highly elastic protein in connective tissue and allows many tissues in the body to recover their shape after stretching or contracting. The FIRELISA Human Elastin ELISA kit assay was designed to measure elastin in tissue or protein / peptide compositions.

[0354] Human dermal fibroblasts (HDF) were seeded at 50,000 cells / well in a 24 well plate in DMEM containing 10% foetal calf serum (FCS), 1% penicillin / streptomycin (Pen / strep), 1% L-glutamine and left for 24 hours.

[0355] After the initial 24 hour incubation, the cells were incubated with 5 μg / ml, 1 μg / ml or 0.1 μg / ml of synthetic peptide for 96 hours respectively.

[0356] After treatment, cells were lysed using 4 freeze-thaw cycles in liquid nitrogen. Lysed cells were centrifuged and 50 μl / ml of each supernatant was assayed using the FIRELISA Human Elastin ELISA kit. All steps were performed according to the manufacturer's instructions.

[0357] Results were calculated as a percentage of the untreated control group. An increase in the optical density reading indicates an increase in collagen content.

[0358] Results are shown in Figure 101 , 103 , 105, 107, 108 and 109.

[0359] Example 4 - Elastin and Cell Proliferation Assays

[0360] Table 2. Test items. Orange bands correspond to samples dissolved in 0.3% DMSO instead of water.

[0361]

[0362] Equipment

[0363] Incubator, flow laminar chamber, sterile polished plastic stick, pipette, maintenance medium, 6-well plate, 24-well plate.

[0364] Reagents

[0365] MTT, PBS, SDS, formaldehyde, dimethylbenzene, absolute ethanol, Dulbecco's Phosphate Buffer Saline (DPBS), Metal Enhanced DAB substrate kit, ABC Peroxidase Staining Kit, citric acid, 2N sodium hydroxide, 30% hydrogen peroxide, Anti-Filaggrin, Anti-rabbit IgG-Biotin, Tween 20.

[0366] Test system

[0367] Nature: human skin tissue 5 mm diameter

[0368] Batch number: EXP004050B009 and EXP004050B011

[0369] Supplier: Laboratoire Biopredic International - 8-18 rue Jean Pecker - 35000 Rennes France. Phone: +33 (0)2.99.14.36.14 - Fax: +33 (0)2.99.54.44.72.

[0370] Analysis certificate in annex 1.

[0371] Two batches were used for the assay. Batch EXP004050B005 was used for the first day of the experiment and batch EXP004050B006 was used for the fifth day of the experiment.

[0372] Maintenance medium

[0373] Maintenance medium: batch number: MIL 218C

[0374] Supplier: Biopredic International Laboratory - 8-18 rue du Clos Court - 35000 Rennes - France Peptide P1 tested: SEQ ID NO: 283

[0375] P2: SEQ ID NO: 246

[0376] P3: SEQ ID NO: 284

[0377] P4: RPYYSNAPQEIF

[0378] P5: VLLEQQEQEPQH

[0379] P6: SEQ ID NO: 245

[0380] P7: QQYGIAASPFLQSAA

[0381] P8: SEQ ID NO: 42

[0382] Composition tested

[0383] P9 (14-CHL-0723-09) is a Pea composition (SEQ ID NO: 50, 85, 74, 140, 82, 136, 189, 77, 169, 149, 171, 178, 143, 127, 190, 141, 147, 133, 186, 125, 122, 119, 87, 90, 86, 89, 138, 129, 123, 120, 117, 113, 110, 121, 105, 98, 55, 161, 19, 317, 135, 130, 146, 177, 160, 170, 188, 83, 78, 36, 96, 159, 26, 330, 168, 148, 184, 151, 151, 165, 114, 284)

[0384] P10 (14-CHL-0723-010) is a Rice composition (SEQ ID NO: 245, 246, 263, 250, 257, 259, 276, 255, 251, 264, 256, 266, 274, 270, 269, 356, 245, 380, 262, 258, 356, 218, 252, 358, 271, 253, 344, 275, 272, 226, 224, 220, 248, 261, 265, 373, 375, 247, 249, 363, 273, 343, 273, 362)

[0385] Method of application

[0386] Skin explants were prepared from abdominoplasty surgery. Some explants were defatted with alcohol to obtain dehydrated skin.

[0387] These explants were cultured for 5 days in a maintenance medium supplied by the supplier, Biopredic International. The test item was applied twice daily at 5 μL per explant.

[0388] At the end of the assay, viability controls were performed using MTT on two explants and the third explant was fixed in 4% formaldehyde for histological analysis and cell staining.

[0389] For each analysis (D1 and D5), histological analysis was performed on defatted explants, explants treated with the test item, DMSO 0.3% control group and water control group.

[0390] Each skin explant in maintenance medium was defatted with 5 μL of alcohol during 3 hours after receipt in the laboratory.

[0391] After 3 hours, all skin explants were treated with the test item twice a day and incubated at 37°C ± 2°C, 5% CO2 for 1 or 5 days.

[0392] Viability controls with MTT were used on days 1 and 5 to ensure system integrity.

[0393] Immunostaining

[0394] Histology was performed by the laboratory Gredeco, and immunostaining for elastin and Ki67 was performed by the same laboratory. Immunostaining for filaggrin was performed by the laboratory Intertek.

[0395] Elastin (rabbit monoclonal antibody, clone P 15502, LSBio) was detected using two layers of immunoperoxidase technology (ABC kit, Vector Laboratories) and visualized by AEC (3-amino-9-ethylcarbazole). The intensity of immunohistochemical staining in elastic fibers was assessed using a semi-quantitative histological score.

[0396] Epithelial cell proliferation was analyzed by immunohistochemistry using anti-Ki67 antibodies. Immunodetection was performed using three layers of indirect immunoperoxidase technique and amplified (DAKO kit) and displayed by AEC (3-amino-9-ethylcarbazole). The number of labeled cells (keratinocytes in the basal layer of the epidermis) was counted and the total number of basal cells was provided to calculate the percentage of labeled cells.

[0397] Specific staining for filaggrin was performed using immunoperoxidase staining (ABC kit, Fisher).The intensity of immunohistochemical labeling in the epidermis was assessed relative to a negative control of solvent (water or DMSO 0.3%).

[0398] C: Results

[0399] Vitality control

[0400] Figure 1 Integrity and viability controls are shown. These controls do allow for validation of the assay system. The viability of the tested items was >50% and, according to the test, they did not show cytotoxicity.

[0401] Immunostaining

[0402] Elastin expression

[0403] The elastic fibers of the dermis were visualized by staining with catechins and morphological quantification was performed by computer-assisted image analysis. The area percentage occupied by elastic fibers in the dermis was calculated in the dermis and the average superficial dermis. The results are shown in Table 1. Figure 2 and Figure 3 middle.

[0404] Table 3. Morphological quantification of elastic fibers in the superficial dermis (%) and the middle dermis. The orange band corresponds to samples dissolved in 0.3% DMSO but not in water.

[0405]

[0406] Under the experimental conditions of this study, samples 0723-1 and 0723-3 showed a two-fold increase in elastic fibers in the superficial dermis compared to the water control group (Figure), and an increase in the middle dermis compared to the water control group at D5.

[0407] Compared to the water control group, the 0723-2 sample showed a two-fold increase in the mid-dermis at day 1 and an increase at day 5.

[0408] Ki67 expression

[0409] The results of the immunohistochemical analysis of Ki67 are reported in the table and expressed as the percentage of labeling at the basal layer of the epidermis. The figure shows the percentage of Ki67 cells compared to the negative control (water or DMSO). The immunohistochemical analysis of mitotic activity is shown in Table 4, with the average value of each analytical condition.

[0410] Table 4. Percentage of Ki67-positive cells in the basal layer of the epidermis. The orange band corresponds to samples dissolved in 0.3% DMSO but not in water.

[0411]

[0412] Under the experimental conditions of the study, test items 0723-06, 0723-08, 0723-09 and 0723-010 showed an increase in the number of mitotic cells compared to EGF at day 1. For all the analyzed conditions, a decrease in the mitotic index was observed at day 5 compared to day 1.

[0413] This decrease in cell staining at day 5 is due to the model. Indeed, after about 3 days, the cell renewal on this model stops.

[0414] The present application is not limited to the embodiments described previously, and can be modified in structure and details without departing from the spirit of the application.

[0415]

[0416]

[0417]

[0418]

[0419]

[0420]

[0421]

[0422]

[0423]

[0424]

[0425]

[0426]

[0427]

[0428]

[0429]

[0430]

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[0437]

Claims

1. A peptide having the amino acid sequence of SEQ ID NO: 283, or a biologically active variant of SEQ ID NO: 283, wherein the variant is selected from GPVEMPYTLLYPSSK, HGPVEMPYTLLYPS, HGPVEMPYTLLYPSS and PVEMPYTLLYPS, or an amino acid sequence of a biologically active variant of SEQ ID NO: 245, wherein the variant is selected from FGKTVFDGVLRPGQL, GKTVFDGVLRPGQL and DGVLRPGQLL.

2. The peptide according to claim 1, wherein the amino acid sequence of the peptide is SEQ ID NO:

283.

3. The peptide according to any one of the preceding claims, which is chemically modified, wherein the peptide is skin growth promoting.

4. The peptide according to claim 3, wherein the peptide is chemically modified by modifying side chains, incorporating protecting groups during peptide synthesis, incorporating non-natural amino acids and / or their derivatives, and using cross-linking agents and methods of imposing conformational constraints on the peptide.

5. A conjugate comprising a peptide according to any one of the preceding claims conjugated, linked or fused to a binding ligand, wherein the binding ligand is selected from a polyethylene glycol polymer, a lipophilic group or an antibody molecule. 6 . A composition comprising the peptide according to claim 1 or the conjugate according to claim 5 .

7. The composition of claim 6, wherein the composition is a powder.

8. The composition of claim 6, wherein the composition is formulated for topical application to human skin.

9. The composition according to claim 8, which is a preparation selected from the group consisting of creams, aqueous dispersions, oils, foams, gels, liniments, pastes, ointments, powders, sprays, aerosols, solutions, suspensions, emulsions, syrups, elixirs, polysaccharide films, patches and bandages.

10. The composition according to claim 9, wherein the emulsion is selected from the group consisting of a multiple emulsion, a water-in-oil emulsion, an oil-in-water emulsion, and a silicone emulsion.

11. The composition of claim 9, wherein the solution is selected from the group consisting of a hydroalcoholic solution and a hydroglycolic acid solution.

12. The composition of claim 9, wherein the gel is selected from a hydrogel.

13. The composition according to claim 9, wherein the patch is selected from a gel patch.

14. The composition according to any one of claims 6 to 13, further comprising a cosmetically or pharmaceutically acceptable excipient selected from the group consisting of diluents, binders, lubricants, suspending agents, coating agents, preservatives, stabilizers, solubilizers, emollients, emulsifiers, humectants and surfactants.

15. The composition according to any one of claims 6 to 13, which is a cosmetic composition.

16. A pharmaceutical composition comprising the peptide according to any one of claims 1 to 4, or the conjugate according to claim 5.

17. A personal care composition comprising the peptide of any one of claims 1 to 4, or the conjugate of claim 5.

18. A wound dressing comprising the peptide of any one of claims 1 to 4, or the conjugate of claim 5, or the composition of any one of claims 6 to 15.

19. The wound dressing of claim 18, wherein the dressing is selected from a plaster, a bandage or a dressing.

20. Use of the peptide according to any one of claims 1 to 4, or the conjugate according to claim 5, or the composition according to any one of claims 6 to 15 in the preparation of a medicament for slowing down or inhibiting aging of human skin.

21. Use of a peptide having an amino acid sequence of SEQ ID NO: 245 in the preparation of a medicament for slowing down or inhibiting human skin aging.

22. Cosmetic use of a peptide according to any one of claims 1 to 4, or a conjugate according to claim 5, or a composition according to any one of claims 6 to 15 for slowing or inhibiting aging of human skin or for treating or caring for visible signs of aging.

23. The cosmetic use according to claim 22, wherein the visible signs of aging are wrinkles.

24. Cosmetic use of a peptide having the amino acid sequence SEQ ID NO: 245 for slowing or inhibiting aging of human skin or for treating or caring for visible signs of aging.

25. The cosmetic use according to claim 24, wherein the visible signs of aging are wrinkles.

26. The cosmetic use according to any one of claims 22 to 25, wherein the peptide is administered topically.

27. Use of the peptide of any one of claims 1 to 4, or the conjugate of claim 5, or the composition of any one of claims 6 to 15 in the preparation of a medicament for treating a wound in a mammal.

28. Use of a peptide having the amino acid sequence SEQ ID NO: 245 in the preparation of a medicament for treating wounds in a mammal.

29. The use according to claim 20, 21, 27 or 28, wherein the drug is a drug for topical administration.

30. A wound dressing comprising a peptide having an amino acid sequence of SEQ ID NO: 245, wherein the dressing is selected from a plaster, a bandage or a dressing.

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